Compounds having reversible inhibiting activity of carnitine palmitoyl-transferase

ABSTRACT

Compounds of formula (I)                    
     wherein the groups are as defined in the description are disclosed. 
     The compounds of formula (I) are endowed with reversible inhibiting activity of carnitine palmitoyl-transferase and are useful in the preparation of medicaments useful in the pathologies related to a hyperactivity of carnitine palmitoyl-transferase, such as hyperglycemia, diabetes and pathologies related thereto, heart failure, ischemia.

This application is a continuation of PCT/IT00126 filed May 11, 1999.

The present invention relates to compounds having inhibiting activityagainst carnitine palmitoyl transferase. The present invention relatesalso to pharmaceutical compositions containing at least one of thesecompounds active ingredients and to the use of said compounds in thepreparation of medicaments useful in the treatment of pathologiesrelated to a hyperactivity of carnitine palmitoyl-transferase, inparticular hyperglycaemic states, such as diabetes and relatedpathologies and of congestive heart failure.

BACKGROUND OF THE INVENTION

To date, hypoglycaemic therapy is based on the use of drugs havingdifferent mechanism of action (Arch. Intern. Med., 1997, 157,1802-1817).

Insulin and its analogues represent the most used therapy, recurring tothe direct hypoglycaemic action of this hormone.

Other compounds act indirectly by stimulating insulin release(sulphonylureas). A different target of hypoglycaemic drugs isrepresented by the reduction of glucose intestinal absorption throughthe inhibition of intestinal glucosidases, or by reducing insulinresistance.

Hyperglycaemia is also treated with gluconeogenesis inhibitors, such asbiguanides.

Some words have also stressed out the relationship betweengluconeogenesis and fatty acid oxidation.

The membrane bound long-chain acylcarnitine transferases, also known ascarnitine palmitoyltransferase (CPT), are widely represented in organsand subcellular organelles (Bieber, L. L. 1988 Ann. Rev. Biochem. 57:261-83). The well-established role of this category of enzymes is thetransport of activated long-chain fatty acids through mitochondrialmembranes. In this context, the outer mitochondrial membrane CIT Icatalyzes the formation of long-chain acylcarnitines that aretransported across the mitochondrial membrane by a specific carrier, andreconverted into long-chain acyl-coenzyme A esters by CPT II, whichresides in the inner mitochondrial membrane. Long-chain acyl-CoAs arethen oxidised to acetyl-coenzyme A, which activates a keygluconeogenetic enzyme: pyruvate carboxylase.

Other works report that diabetic patients have high blood levels offatty acids, whose liver oxidative fate gives rise to an increase ofacetyl-coenzyme A, ATP and NADH. High availability of these compoundsmaximally stimulates gluconeogenesis, which is in part responsible ofthe elevated glucose blood levels in diabetic patients. CPT inhibitionindirectly reduces the extent of liver gluconeogenesis, and hence bloodglucose levels.

CPT inhibitors have been disclosed in J. Med. Chem., 1995, 38(18),3448-50 and in the corresponding European patent application EP 0 574355 as potential derivatives with hypoglycaemic activity.

Aminocarnitines N-acylated with —COR residue, wherein R is an aliphaticresidue with 1 to 19 carbon atoms are disclosed in WO85/04396 useful forinvestigating the role of transferases in the body, in particular thespecificity of carnitine acyltransferase.

Emeriamine and its analogues are disclosed in EP 0 127 098 and J.Med.Chem. 1987, 30, 1458-1463.

Notwithstanding the mechanism of activity above outlined, to date, drugsinhibiting CPT capable to effectively counteract hyperglycaemia do notexist. For some products, such as tetradecyl glycidic acid, or etomoxir,myocardial hypertrophy have been evidenced as side effects (Life Sci.,1989, 44, 1897-1906).

None of the therapies presently used in clinic is fully satisfying, inparticular due to the onset of unwanted side effects, such as severehypoglycaemia, allergic phenomena, oedema, diarrhoea, intestinaldisturbances, kidney toxicity, etc.

The necessity to obtain alternative effective therapies forhyperglycaemia still remains.

ABSTRACT OF THE INVENTION

It has now surprisingly been found that compounds of general formula(I):

wherein: X⁺ is selected from the group consisting of N⁺(R₁,R₂,R₃) andP⁺(R₁,R₂,R₃), wherein

(R₁,R₂,R₃), being the same or different, are selected in the groupconsisting of hydrogen and C₁-C₉ straight or branched alkyl groups,—CH═NH(NH₂), —NH₂, —OH; or two or more R₁, R₂ and R₃ together with thenitrogen atom, which they are linked to, form a saturated orunsaturated, monocyclic or bicyclic heterocyclic system; with theproviso that at least one of the R₁, R₂ and R₃ is different fromhydrogen;

Z is selected from

—OR₄,

—OCOOR₄,

—OCONHR₄,

—OCSNHR₄,

—OCSOR₄,

—NHR₄,

—NHCOR₄,

—NHCSR₄,

—NHCOOR₄,

—NHCSOR₄,

—NHCONHR₄,

—NHCSNHR₄,

—NHSOR₄,

—NHSONHR₄,

—NHSO₂R₄,

—NHSO₂NHR₄,

—SR₄,

wherein —R₄ is a C₁-C₂₀ saturated or unsaturated, straight or branchedalkyl group, optionally substituted with a A₁ group, wherein A₁ isselected from the group consisting of halogen atom, C₆-C₁₄ aryl,heteroaryl, aryloxy or heteroaryloxy group, said aryl, heteroaryl,aryloxy or heteroaryloxy groups being optionally substituted with one ormore C₁-C₂₀ saturated or unsaturated, straight or branched alkyl oralkoxy group and/or halogen atom;

Y⁻ is selected from the group consisting of —COO—, PO₃H⁻, —OPO₃H⁻,tetrazolate-5-yl;

with the proviso that when Z is —NHCOR₄, X⁺ is trimethylammonium, Y is—COO—, then R₄ is C₂₀ alkyl;

with the proviso that when Z is —NHSO₂R₄, X⁺ is trimethylammonium and Y⁻is —COO—, then R₄ is not tolyl;

with the proviso that when Z is —NHR₄, X⁺ is trimethylammonium and Y⁻ is—COO—, then R₄ is not C₁-C₆ alkyl.

The present invention further comprises the use of the compounds of theabove-mentioned formula (I) as active ingredients for medicaments, inparticular for medicaments useful for the treatment of pathologiesrelated to a hyperactivity of carnitine palmitoyl carnitine, such as andin particular hyperglycemic states, diabetes and related pathologies,congestive heart failure and dilatative cardiopathy.

The present invention comprises pharmaceutical compositions containingcompounds of formula (I) as active ingredients, in admixture withpharmaceutically acceptable vehicles and excipients.

The present invention comprises also processes for the preparation ofcompounds of formula (I).

DETAILED DESCRIPTION OF THE INVENTION

Within the scope of the present invention, as examples of C₁-C₂₀ linearor branched alkyl group, methyl, ethyl, propyl, butyl, pentyl, hexyl,heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl,pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl and eicosyl andtheir possible isomers are meant, such as for example isopropyl,isobutyl, tert-butyl.

Examples of C₁-C₂₀ linear or branched alkenyl group are methylene,ethylidene, vinyl, allyl, propargyl, butylene, pentylene, wherein thecarbon-carbon double bond, optionally in the presence of othercarbon-carbon unsaturations, can be situated in the different possiblepositions of the alkyl chain, which can also be branched within theallowed isomery.

Examples of (C₆-C₁₄) aryl group are phenyl, 1- or 2-naphthyl, anthryl,optionally substituted as shown in the general definitionsabove-mentioned.

Examples of heterocyclic groups thienyl, quinolyl, pyridyl,N-methylpiperidinyl, 5-tetrazolyl, optionally substituted as shown inthe general definitions above-mentioned.

As halogen atom it is intended fluorine, chlorine, bromine, iodine.

The compounds of formula (I) can be also in the form of inner salts.

A first group of preferred compounds comprises the compounds of formula(I) wherein N⁺ (R₁,R₂,R₃) is trimethyl ammonium.

A second group of preferred compounds comprises the compounds of formula(I) wherein two or more R₁, R₂ and R₃, together with the nitrogen atom,which they are linked to, form a saturated or unsaturated, monocyclic orbicyclic heterocyclic system; for example morpholinium, pyridinium,pyrrolidinium, quinolinium, quinuclidinium.

A third group of preferred compounds comprises the compounds of formula(I) wherein R₁ and R₂ are hydrogen and R₃ is selected from the groupconsisting of —CH═NH(NH₂), —NH₂ and —OH.

Within the different embodiments of the present invention, the R₄ groupis preferably a C₇-C₂₀ saturated or unsaturated, straight or branchedalkyl group. In fact, it has been observed the length of the alkyl chainR₄ significantly increases the selectivity against CPT. referred R₄groups are selected from the group consisting of heptyl, octyl, nonyl,decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl,heptadecyl, octadecyl, nonadecyl and eicosyl.

Preferred examples of Z group are ureido (—NHCONHR₄), and carbamate(—NHCOOR₄, —OCONHR₄) ones.

In particular, compounds of formula (I) wherein X⁺, R₁, R₂, R₃, have theabove disclosed meanings, Z is ureido (—NHCONHR₄) or carbamate(—NHCOOR₄, —OCONHR₄), R₄ is a C₇—C₂₀, preferably a C₉-C₁₈ saturated orunsaturated, straight or branched alkyl group, are preferred.

The compounds of formula (I) have an asymmetry center on carbon atombound to a Z group. For the purposes of the present invention, eachcompound of formula (I) can exist both as R,S racemic mixture and asseparated R/S isomeric form.

The compounds of formula (I) are quaternary ammonium or phosphoniumderivatives always containing a Y⁻ anionic group. Dependently on pH,each compounds of formula (I) can exist indifferently as amphoion (innersalt) or as a compound wherein Y⁻ is present in the YH form. In such acase, X⁺ is salified with a pharmacologically acceptable acid. Formula(I) covers all these different possibilities. In case of nitrogen atomshaving basic character, the salts with pharmaceutically acceptableacids, both inorganic and organic, such as for example, hydrochloricacid, sulfuric acid, acetic acid, or, in the case of acid group, such ascarboxyl, the salts with pharmaceutically acceptable bases, bothinorganic and organic, such as for example, alkaline and alkaline-earthhydroxides, ammonium hydroxide, amine, also heterocyclic ones. Examplesof pharmaceutically acceptable salts are chloride; bromide; iodide;aspartate; acid aspartate; citrate; acid citrate; tartrate; acidtartrate; phosphate, acid phosphate; fumarate; acid fumarate;glycerophosphate; glucosephosphate; lactate; maleate; acid maleate;mucate; orotate; oxalate; acid oxalate; sulfate; acid sulfate;trichloroacetate; trifluoroacetate; methanesulfonate; pamoate and acidpamoate.

A first group of particularly preferred compounds comprises:

R,S-4-trimethylammonium-3-(nonylcarbamoyl)-aminobutyrate;

R,S-4-quinuclidinium-3-(tetradecyloxycarbonyl)-oxybutyrate;

R,S-4-trimethylammonium-3-(nonylcarbamoyl)-oxybutyrate;

R,S-4-trimethylammonium-3-(nonyloxycarbonyl)-oxybutyric acid chloride;

R,S-4-trimethylphosphonium-3-(nonylcarbamoyl)-oxybutyrate;

R,S-4-trimethylammonium-3-(octyloxycarbonyl)-aminobutyrate;

R,S-4-trimethylammonium-3-(nonyloxycarbonyl)-aminobutyrate;

R,S-4-trimethylammonium-3-octyloxybutyrate;

R,S-4-trimethylammonium-3-tetradecyloxybutyrate;

R,S-1-guanidinium-2-tetradecyloxy-3-(tetrazolate-5-yl) -propane;

R,S-1-trimethylammonium-2-tetradecyloxy-3-(tetrazolate-5-yl)-propane;

R,S-3-quinuclidinium-2-(tetradecyloxycarbonyl)-oxy-1-propanephosphonatemonobasic;

R,S-3-trimethylammonium-2-(nonylaminocarbonyl)-oxy-1-propanephosphonatemonobasic;

R,S-3-pyridinium-2-(nonylaminocarbonyl)-oxy-1-propanephosphonic acidchloride;

R-4-trimethylammonium-3-(tetradecylcarbamoyl)-aminobutyrate;

R-4-trimethylammonium-3-(undecylcarbamoyl) -aminobutyrate;

R-4-trimethylammonium-3-(heptylcarbamoyl)-aminobutyrate;

R-4-trimethylammonium-3-(nonylthicarbamoyl)-aminobutyrate;

R,S-4-trimethylammonium-3-(nonylthiocarbamoyl)-aminobutyrate;

R-4-trimethylammonium-3-(nonylcarbamoyl)-aminobutyrate;

S-4-trimethylammonium-3-(nonylcarbamoyl)-aminobutyrate;

S-4-trimethylammonium-3-(tetradecylcarbamoyl)-aminobutyrate;

R,S-4-trimethylammonium-3 -tetradecylaminobutyrate;

R,S-4-trimethylammonium-3-octylaminobutyrate;

R,S-4-trimethylammonium-3-(decansulfonyl)aminobutyrate;

R,S-4-trimethylammonium-3-(nonylsulfamoyl)aminobutyrate;

S-4-trimethylammonium-3-(dodecansulfonyl)aminobutyrate;

R-4-trimethylammonium-3-(dodecansulfonyl)aminobutyrate;

S-4-trimethylammonium-3-(undecylsulfamoyl)aminobutyrate;

R-4-trimethylammonium-3-(undecylsulfamoyl)aminobutyrate;

R-4-trimethylammonium-3-(dodecylcarbamoyl)aninobutyrate;

R-4-trimethylammonium-3-(10-phenoxydecylcarbamoyl)aminobutyrate;

R-4-trimethylammonium-3-(trans-β-styrenesulfonyl)aminobutyrate.

The compounds of formula (I) can be prepared with reactions that arewell known in the state of the art. A process for the preparation of thecompounds of claim 1, wherein Z is —NHR₄ comprising the reaction ofX⁺—CH₂—CH(NH₂)—CH₂—Y⁻, wherein X⁺ and Y⁻ have the same meanings as inclaim 1, of the desired structure, optionally protected on the acid Y⁻group, with alkane carbaldheydes, wherein the alkyl moiety is a one-termlower homologue of the desired R₄, and subsequent reduction.

Generally, the compounds of formula (I), wherein Z is carbonate(—OCOOR₄), carbamate (—OCONHR₄, —NHCOOR₄), thiocarbamate (—OCSNHR₄,—NHCSOR₄,) or thiocarbonate (—OCSOR₄), are obtained by reacting acompound of formula X⁺—CH₂—CH(OH)—CH₂—Y⁻, wherein X⁺ and Y⁻ are as abovedefined, of the desired structure, optionally protected on the acid Y⁻group, respectively with alkyl chloroformates, alkyl isocyanates, alkylisothiocyanates, alkyl thiochloroformates, containing the desired R₄alkyl group.

Compounds of formula (I), wherein Z is amide (—NHCOR₄), thioamide(—NHCSR₄), carbamate (—NHCOOR₄, —OCONHR₄), thiocarbamate (—NHCSOR₄,—OCSNHR₄,), ureido (—NHCONHR₄), thioureido (—NHCSNHR₄), sulfinamide(—NHSOR₄), sulfonamide (—NHSO₂R₄), sulfinamoylamino (—NHSONHR₄), andsulfamide (—NHSO₂NHR₄), are obtained by reacting X⁺—CH₂—CH(NH₂)—CH₂—Y—,wherein X⁺ and Y⁻ are as above defined, of the desired structure,optionally protected on the acid Y⁻ group, respectively with acylchlorides, thioacyl chlorides, alkyl chloroformates, alkylthiochloroformates, alkyl isocyanates, alkyl thioisocyanates, alkylsulfinyl chlorides, alkyl sulfonyl chlorides, SOCl₂ and alkyl amines,alkyl sulfamoyl chlorides (or SO₂Cl₂ and alkyl amines), containing thedesired R₄ alkyl group.

Compounds of formula (I), wherein Z is —OR4 or —SR4 are obtained by thereaction of carbonyl compounds of formula Hal-CH2—CO—CH2—COOR′, whereinHal is a halogen atom, preferably chlorine, and R′ is the residue of asuitable ester, such as for example a lower alkyl ester (an ethyl or atert-butyl ester) with respectively alcohols and thiols R4OH or R4SH,wherein R4 is as above defined, to give the respective ketal orthioketal, followed by the transformation of the respective ketal orthioketal into the respective ether or thioether, subsequentsubstitution of the Hal atom with a nucleophilic group, such as azido,phthalimido, nitro, amino, alkyl amino group, and transformation of thenucleophilic group into the X+ group, wherein X+ is N⁺(R₁,R₂,R₃) or,alternatively the Hal atom is substituted with a (R₁,R₂,R₃)-substitutedphosphine to obtain the compounds of formula (I) wherein X⁺ isP⁺(R₁,R₂,R₃).

Compounds of formula (I), wherein Z is —NHR₄ are obtained by reactingX⁺—CH₂—CH(NH₂)—CH₂—Y⁻, wherein X⁺ and Y⁻ have the same meanings as inclaim 1, of the desired structure, optionally protected on the acid Y⁻group, with alkane carbaldheydes, wherein the alkyl moiety is a one-termlower homologue of R₄ and subsequent reduction.

Regarding the various meanings of R₄, present in the differentreactives, these reactives are available in the market, or can beprepared according to well-known methods in literature, which theexperts in the field can resort to, completing with their own knowledgeof the argument.

Pharmaceutically acceptable salts are obtained with conventional methodsfound in the literature, and do not necessitate of further disclosure.

The compounds disclosed in the present invention have reversibleinhibiting activity of carnitine palmitoyl-transferase (CPT). Thisactivity allows their use as active ingredients in the preparation ofmedicaments useful for the treatment and prevention of hyperglycaemia,diabetes and disorders related thereto, such as, for example diabeticretinopathy, diabetic neuropathy. The compounds of the present inventionare also useful as active ingredient for the treatment and prevention ofcardiovascular disorders, such as congestive heart failure. Thecompounds of formula (I) are also applicable for medicaments for theprevention and treatment of ketonic states, wherein it is intended thepathological conditions characterized by high levels of ketone bodies inblood.

Inhibiting activity mainly occurs on the isoform I of palmitoylcarnitine transferase (CPT-I).

A further object of the present invention relates to pharmaceuticalcompositions comprising at least a compound of formula (I), in an amountsuch as to produce a significant therapeutical effect. The compositionsaccording to the present invention are conventional and are obtainedwith commonly used methods in the pharmaceutical industry. According tothe desired administration route, the compositions shall be in solid orliquid form, suitable to the oral, parenteral, intravenous ortransdermal route. The compositions according to the present inventioncomprise together with the active ingredients at least apharmaceutically acceptable vehicle or excipient. Formulationco-adjuvants, for example solubilizing, dispersing, suspending,emulsionating agents can be particularly useful. Examples of suitableoral pharmaceutical compositions are capsules, tablets, granulates,powders, syrups, elixirs. Examples of suitable parenteral pharmaceuticalcompositions are solutions, emulsions, suspensions. Examples of suitabletransdermal pharmaceutical compositions are patches, subcutaneousimplants.

The compounds of formula (I) can also be used in combination with otherwell-known active ingredients.

The dose of the active ingredients will vary depending on the kind ofactive ingredient used, the administration route, the grade of pathologyto be treated and the general conditions of the subject. The dosage andposology shall be determined by the clinic expert or the physician.Generally, a therapeutic effect can be obtained at dosages comprisedbetween 1 and 100 mg/kg body weight.

The compounds according to the present invention are useful asmedicaments with hypoglycaemic activity. A further object of the presentinvention is the preparation of a pharmaceutical composition comprisingadmixing at least a compound of formula (I) with suitablepharmaceutically acceptable excipients and/ or vehicles.

The following examples further illustrate the invention.

EXAMPLE 1 R,S-4-trimethylammonium-3-(nonylcarbamoyl)-aminobutyrate (ST1251)

Nonyl Isocyanate

A solution of decanoyl chloride (20 g, 104.8 mmoles) in acetone (30 ml)was dropped into a solution of sodium azide (9.53 g, 146.6 mmoles) inwater (30 ml), cooled in an ice bath. The temperature of the azidesolution was kept between 10 and 15° C. after one hour, the solution wastransferred in a separatory funnel and the lower phase (the aqueous one)was eliminated. The higher phase was transferred into a flask containing100 ml of toluene, previously warmed at 65° C. After 1.5 hours, thesolution was evaporated to dryness, giving 13.37 g crude product, whichafter vacuum distillation gave 8.3 g pure product in the form ofcolourless liquid.

Yield 47%. ¹H-NMR (300 MHz; CDCl₃): δ: 3.3 (t, 2H), 1.6 (m, 2H),1.45-1.2 (m, 12H), 0.9 (brt, 3H).

R,S-4-trimethylammonium-3-(nonylcarbamoyl)-aminobutyrate

Nonyl isocyanate (15.42 g, 91.12 mmoles) was added to a solution ofaminocarnitine, inner salt (7.3 g, 45.56 mmoles) in anhydrous DMSO (350ml) and the solution was left to stand for 60 hours at 40° C. Theresulting mixture was transferred in a 3 l Erlenmeyer flask, containingethyl ether (2.5 l) and the solvent was separated by decanting theformed precipitate, which was then transferred into a flask andprecipitated again with ethyl ether. The so obtained crude product waswashed several times with ethyl ether and purified on a silica gelchromatographic column, using a CHCl₃: MeOH 9:1 to CHCl₃: MeOH 3:7gradient until elution of impurities with higher Rf, then eluting theproduct of interest with MeOH only. 9.7 g of pure product were obtained.

Yield 68%. M.p.: 145-147° C. ¹H-NMR (300 MHz; D₂O): δ: 4.4 (m, 1H), 3.45(dd, 1H), 3.30 (d, 1H), 3.05 (s, 9H), 2.9 (t, 2H), 2.3 (d, 2H), 1.3 (m,2H), 1.15 (brs, 12H), 0.8 (brt, 3H). FAB Mass=330, [(M+H)⁺]. Elementalanalysis: responding to the expected formula C₁₇H₃₅N₃O₃. K.F.=2.5%water. TLC silica gel CHCl₃: iPrOH:MeOH:H₂O:CH₃COOH 42:7:28:10.5:10.5;Rf=0.55. HPLC: SGE-SCX column (5 μm, 250×4 mm), T=30° C., mobile phase0.2 M KH₂PO₄:CH₃CN 85:15, pH as such, flow 0.75 ml/min, detector: RI, UV205 nm, RT=12.63 min.

EXAMPLE 2 R,S-4-quinuclidinium-3-(tetradecyloxycarbonyl)-oxybutyrate (ST1265)

ter-Butyl R,S-4-guinuclidinium-3-hydroxybutyrate Iodide

Quinuclidine (2.40 g, 21.60 mmoles) was added to ter-ButylR,S-4-iodo-3-hydroxybutyrate (6.18 g, 21.60 mmoles) in acetonitrile (60ml) and the solution was warmed to 60° C. for 20 hours under stirring.After evaporation of the solvent, the residue was dissolved inacetonitrile and precipitated with ethyl ether several times to give 7.2g of product, contaminated with about 13% by weight of quinuclidineiodide (as from NMR). After repeated crystallization from CH₃CN/Et₂O,4.3 g of pure product were obtained.

Yield 50%. M.p.: 124-127° C. ¹H-NMR (300 MHz; D₂O): δ: 4.50 (m, 1H),3.40 (m, 2H), 2.42 (m, 2H), 2.08 (m, 1H), 1.88 (m, 6H), 1.34 (m, 9H).FAB Mass=270, [M⁺]. Elemental analysis: responding to the expectedformula C₁₅H₂₈ INO₃. K.F.=0.5% water. The preparation of ter-butyl4-iodo-3-hydroxybutyrate was carried out as described in J. Pharm.Science 64/7, 1262-1264, 1975.

Tetradecyl Chloroformate

29 ml of a 20% toluene solution of phosgene (55.98 mmoles) was added totetradecyl alcohol (4 g, 18.66 mmoles) and the reaction mixture was leftto stand for 20 hours under stirring at room temperature. After solventevaporation, the residue was taken up with hexane and evaporated todryness (several times) to give 5.1 g product as colourless liquid.

Yield 98%. ¹H-NMR (300 MHz; CDCl₃): δ: 4.30 (t, 2H), 1.72 (m, 2H), 1.30(m, 22H), 0.85 (brt, 3H).

ter-butyl R,S-4-guinuclidinium-3-(tetradecyloxycarbonyl)-oxy butyratechloride

Dimethylaminopyridine (922 mg, 755 mmoles) and tetradecyl chloroformate(2.09 g, 7.55 mmoles) were added to ter-butylR,S-4-quinuclidinium-3-hydroxybutyrate (2 g, 5.03 mmoles) in anhydrousCH₂Cl₂ (20 ml). The solution was left to stand at room temperature for20 hours under stirring. After this time, the solution was diluted withCHCl₃, saturated with NaCl, and dried over anhydrous sodium sulfate. Thedry residue obtained after evaporation was taken up with ethyl ether andthe undissolved residue was filtered off. After solvent evaporation acrude product was obtained. Flash-chromatography (CHCl₃: MeOH 9:1) andelution with MeOH on Amberlyst A-21 resin (activated in HCl from), gave1.6 g product as chloride.

Yield 58%. M.p.: 59-60° C. ¹H-NMR (300 MHz; CDCl₃): δ: 5.50 (m, 1H),4.55 (d, 2H), 3.80 (m, 7H), 2.90 (dd, 1H), 2.75 (dd, 1H), 2.22 (m, 1H),2.05 (d, 6H), 1.65 (m, 2H), 1.41 (s, 9H), 1.25 (m, 22H), 0.85 (brt, 3H).FAB Mass=510, [M⁺]. Elemental analysis: responding to the expectedformula C₃₀H₅₆ ClNO₅. K.F.=1.5% water.

R,S-4-guinuclidinium-3-(tetradecyloxycarbonyl)-oxybutyrate

Trifluoroacetic acid (6 ml) was added to ter-butylR,S-4-quinuclidinium-3-(tetradecyloxycarbonyl)-oxybutyrate chloride(1.05 g, 1.92 mmoles) and the solution was left to stand for 1 hour atroom temperature under stirring. After vacuum-evaporation oftrifluoroacetic acid, the residue was taken up with cyclohexane andevaporated to dryness several times, then transferred on an AmberlystIRA 402 resin (Cl⁻ form) and eluted with water. The crude product,obtained by freeze-drying was purified through silica gelflash-chromatography (CHCl₃: MeOH 8:2) giving 480 mg product as innersalt.

Yield 55%. M.p.: 132-134° C. ¹H-NMR (300 MHz; D₂O): δ: 5.35 (m, 1H),4.05 (m, 2H), 3.40 (m, 8H), 2.55 (dd, 1H), 2.35 (dd, 1H), 2.08 (m, 1H),1.90 (m, 6H), 1.55 (m, 2H), 1.20 (m, 22H), 0.75 (brt, 3H). FAB Mass=454,[(M+H)⁺. Elemental analysis: responding to the expected formulaC₂₆H₄₇NO₅ K.F.=1.5% water. TLC silica gel CHCl₃:MeOH 7:3. Rf=0.34. HPLC:SGE-SCX column (5 μm, 250×4 mm), T=30° C., mobile phase 0.05 M(NH₄)H₂PO₄:CH₃CN 60:40, pH 4.0, flow 0.75 ml/min, detector: RI, UV 205nm, RT=6.72 min.

EXAMPLE 3 R,S-4-trimethylammonium-3-(nonylcarbamoyl)-oxybutyrate (ST1298)

Benzyl ester of R,S-4-trimethylammonium-3-(nonylcarbamoyl)-oxybutyricacid perchlorate

Nonyl isocyanate (7.39 g, 43.36 mmoles) was added to a solution ofR,S-carnitine perchlorate, benzyl ester (7.69 g, 21.86 mmoles) intoluene (100 ml) and the solution was refluxed for 5 days understirring. Nonyl isocyanate (1.84 g, 10.86 mmoles) was further added andthe reaction mixture was left under reflux for other 5 days. The solventwas vacuum-evaporated and the residue was washed with ethyl ether andsubsequently taken up with chloroform, washed with water and dried overanhydrous sodium sulfate. The oil resulting from the evaporation of theorganic phase was purified through flash-chromatography column, using agradient CHCl₃ to CHCl₃: MeOH 95:5. 4.4 g product were obtained in theform of a thick oil.

Yield 38.6%. ¹H-NMR (200 MHz; CDCl₃): δ: 7.3 (s, 5H), 5.4 (m, 2H), 5.05(m, 2H), 3.8 (dd, 1H), 3.55 (d, 1H), 3.15 (s, 9H), 3.05 (m, 2H), 2.75(m, 2H), 1.4 (m, 2H), 1.2 (brs, 12H), 0.8 (brt, 3H). TLC silica gelCHCl₃: MeOH 9:1; Rf=0.29.

R,S-4-trimethylammonium-3-(nonylcarbamoyl)-oxybutyrate

10% Pd/C (0.44 g) was added to benzyl ester ofR,S-4-trimethylammonium-3-(nonylcarbamoyl)-oxybutyric acid perchlorate(4.4 g, 8.44 mmoles) in MeOH (115 ml) and the mixture was hydrogenatedat 47 psi for 4 hours. After filtration on celite, the solution wasvacuum-concentrated and passed through an Amberlyst A-21 resin, elutingwith MeOH. After solvent evaporation, 2.47 g product were obtained.

Yield 88.7%. M.p.: 151-153° C. ¹H-NMR (300 MHz; D₂O): δ: 5.4 (m, 1H),3.75 (dd, 1H), 3.5 (d, 1H), 3.15 (s, 9H), 3.05 (t, 2H), 2.55 (dd, 1H),2.40 (dd, 1H), 1.45 (m, 2H), 1.20 (brs, 12H), 0.8 (brt, 3H). FABMass=331, [(M+H)⁺]. Elemental analysis: responding to the expectedformula C₁₇H₃₄ N₂O₄. K.F.=1.5% water. TLC silica gel MeOH. Rf=0.22.HPLC: SPHERISORB-SCX column (5 μm, 250×4 mm), T=35° C., mobile phase 50mM KH₂PO₄:CH₃CN 40:60, pH 4.0 with H₃PO₄, flow 0.75 ml/min, detector:RI, UV 205 nm, RT=5.33 min.

EXAMPLE 4 R,S-4-trimethylammonium-3-(nonyloxycarbonyl)-oxybutyratechloride (ST 1297)

Benzyl ester of R,S-4-trimethylammonium-3-(nonylcarbamoyl)-oxybutyricAcid Chloride

Dimethylaminopyridine (3.8 g, 31.2 mmoles) and nonyl chloroformate (6.45g, 31.2 mmoles) were added to R,S-carnitine perchlorate, benzyl ester(7.33 g, 20.8 mmoles) in anhydrous DMF (50 ml) at 0° C. The temperaturewas left to raise to room temperature and the reaction mixture was leftto stand for 3 days under stirring. CHCl₃ was added and the solution waswashed with 1N perchloric acid. The organic phase was dried overanhydrous sodium sulfate and evaporated to dryness, to give 6.02 g crudeproduct, which was purified through flash-chromatography (CHCl₃: MeOH85:15). 3.52 g a thick oil were obtained, which were subsequentlydissolved in MeOH and passed through an Amberlyst A-21 resin (activatedin HCl from), eluting with MeOH. After vacuum-evaporation of thesolvent, 3.1 g oily product were obtained.

Yield 32.4%. ¹H-NMR (200 MHz; CDCl₃): δ: 7.3 (s, 5H), 5.45 (m, 1H), 5.05(s, 2H), 4.4 (d, 1H), 4.1 (t, 2H), 3.8 (dd, 1H), 3.4 (s, 9H), 2.9 (m,2H), 1.55 (m, 2H), 1.2 (brs, 12H), 0.8 (brt, 3H).

Mutatis mutandis, the preparation of nonyl chloroformate was carried outas disclosed in Example 2 for tetradecyl chloroformate.

R,S-4-trimethylammonium-3-(nonyloxycarbonyl)-oxybutyric Acid Chloride

10% Pd/C (110 mg) was added to benzylR,S-4-trimethylammonium-3-(nonyloxycarbonyl)-oxybutyric acid chloride(1.1 g, 2.4 mmoles) in MeOH (10 ml) and the mixture was hydrogenated at47 psi for 2 hours. After filtration on celite, the solution wasvacuum-dried giving 883 mg product (yield 100%), which was furtherpurified by precipitation from CH₃CN/Et₂O. 600 g of product wereobtained.

Yield: 68%. M.p.: 150° C. dec. ¹H-NMR (300 MHz; D₂O): δ: 5.4 (m, 1H),4.1 (m, 2H), 3.75 (dd, 1H), 3.55 (d, 1H), 3.1 (s, 9H), 2.7 (m, 2H), 1.5(m, 2H), 1.20 (brs, 12H), 0.7 (brt, 3H). FAB Mass=332, [M⁺]. Elementalanalysis: responding to the expected formula C₁₇H₃₄ ClNO₅. K.F.=1.7%water. TLC silica gel CHCl₃:MeOH 1:1; Rf=0.10. HPLC: SPHERISORB-C1column (5 μm, 250×4.6 mm), T=30° C., mobile phase 50 mM (NH₄)H₂PO₄:CH₃CN60:40, pH 3.0 with H₃PO₄, flow 0.75 ml/min, detector: RI, UV 205 nm,RT=5.67 min.

EXAMPLE 5 R,S-4-trimethylphosphonium-3-(nonylcarbamoyl)-oxybutyrate (ST1300)

Ethyl ester of R,S-4-trimethylphosphonium-3-hydroxybutyric Acid Iodide

A 1M solution of trimethylphosphine in THF (93 ml) was added to ethylR,S-4-iodo-3-hydroxybutyrate (20 g, 77.5 mmoles) and the reactionmixture was left to stand at room temperature for 5 days under stirring.Ethyl ether was added, and the precipitate formed was separated bydecantation. The precipitate was triturated with Et₂O and dried undervacuum, giving 18.5 g product.

Yield 71.3%. M.p.: 105-107° C. ¹H-NMR (200 MHz; CDCl₃): δ: 4.6 (m, 1H),4.15 (q, 2H), 3.1 (m, 1H), 2.75 (m, 3H), 2.2 (d, 9H), 1.3 (t, 3H).

The ethyl ester of R,S-4-trimethylphosphonium-3-hydroxybutyric acid wasprepared as described in Tetrahedron 1990, 4277-4282, starting fromR,S-3-hydroxy-4-butyrolactone.

Ethyl ester of R,S-4-trimethylphosphonium-3-(nonylcarbamoyl) -oxybutyricAcid Iodide

Nonyl isocyanate (4.04 g, 23.86 mmoles) was added to the ethyl ester ofR,S-4-trimethylphosphonium-3-hydroxybutyric acid iodide (4 g, 11.97mmoles) in anhydrous DMF (80 ml) and the solution was left to stand for7 days at 110° C. under stirring. CHCl₃ was added (300 ml) and thesolution was washed with water and dried over Na₂SO₄. The residueobtained after evaporation of the solvent was taken up withacetonitrile, the formed solid was filtered off and the filtrate waspurified by silica gel flash-chromatography, using CHCl₃: MeOH 8:2. 2.07g of product in the form of a thick oil were obtained.

Yield 34.3%. ¹H-NMR (200 MHz; CDCl₃): δ: 5.4 (m, 2H), 4.15 (q, 2H), 3.15(m, 4H), 2.8 (d, 2H), 2.2 (d, 9H), 1.5 (m, 2H), 1.2 (brs, 12H), 0.8(brt, 3H).

R,S-4-trimethylphosphonium-3-(nonylcarbamoyl) -oxybutyrate Ethyl esterof R,S-4-trimethylphosphonium-3-(nonylcarbarmoyl)-oxybutyric acid iodide(2.07 g, 4.11 mmoles) was dissolved into 1N HCl (200 ml) and thesolution was warmed to 70° C. for 3 hours. The residue obtained aftersolvent vacuum-evaporation was taken up with MeOH and passed throughAmberlyst A-21 resin, eluting with MeOH. A crude product was obtained,which was purified by flash-chromatography, eluting with MeOH and giving700 mg product.

Yield: 49%. M.p.: 123-127° C. dec. ¹H-NMR (300 MHz; D₂O): δ: 5.3 (m,1H), 3.1 (m, 2H), 2.80-2.45 (m, 4H), 1.85 (d, 9H), 1.4 (m, 2H), 1.2(brs, 12H), 0.8 (brt, 3H). FAB Mass=348, [(M+H)⁺]. Elemental analysis:responding to the expected formula C₁₇H₃₄ NO₄P. K.F.=3.4% water. TLCsilica gel MeOH; Rf=0.18.

HPLC: SPHERISORB-SCX column (5 μm, 250×4 mm), T=25° C., mobile phase 50mM KH₂PO₄:CH₃CN 40:60, pH 4.0 with H₃PO₄, flow 0.75 ml/min, detector:RI, UV 205 nm, RT=5.18 min.

The following Examples 6 and 7 are further illustrated by FIG. 1.

EXAMPLE 6 R,S-4-trimethylammonium-3-(octyloxycarbonyl)-aminobutyratechloride (ST 1253) (2a, FIG. 1)

Step A

3 g (0.012 mmoles) aminocarnitine isobutyl ester were dissolved into 20ml anhydrous CH₂Cl₂. 2.48 ml (0.1078 moles) triethylamine and 3.6 g(0.0178 moles) octyl chloroformate (previously prepared by reacting thealcohol with a toluene solution of phosgene) were added to the solution.The reaction mixture was left to stand for 4.5 hours at roomtemperature. Then the solvent was evaporated off and the resulting solidwas dissolved into ethyl acetate and filtered. The solvent wasvacuum-evaporated to dryness and the resulting solid was purified onsilica gel, eluting with 100% CHCl₃, then with CHCl₃:MeOH 95:5 and90:10. The product was obtained with a 50% yield.

TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol 7/water 10.5/aceticacid 10.5)/acetone 7:3; Rf=0.8. HPLC: SPHERISORB-SCX column (5 μm, 250×4mm), mobile phase 50 mM (NH₄)H₂PO₄:CH₃CN 60:40, pH 4.0, detector: RI, UV205 nm, RT=8.6 min. ¹H-NMR (300 MHz; CD₃OD): δ: 4.56-4.46 (m, 1H),4.12-4.02 (m, 2H), 3.94-3.88 (m, 2H), 3.66-3.5 (s, 9H), 3.4 (s, 9H),2.74-2.66 (m, 2H), 2-1.86 (m, 1H), 1.68-1.56 (t, 2H), 1.4-1.2 (m, 12H),0.97-0.7 (d, 6H), 0.6-0.3 (t, 3H). Elemental analysis: responding to theexpected formula C₂₀H₄₁ N₂O₄Cl.

Step B

The ester obtained in step A was hydrolysed on Amberlyst IRA 402 resin(OH⁻ activated form) eluting with water. Water was evaporated todryness; the resulting solid was triturated with acetone andsubsequently filtered. A white solid was obtained.

Yield 94%. M.p.=170° C. dec. ¹H-NMR (300 MHz; CD₃OD): δ: 4.4 (m, 1H),4.05 (t, 2H), 3.5 (d, 2H), 3.2 (s, 9H), 2.4 (d, 2H), 1.6 (m, 2H),1.4-1.2 (m, 12H), 0.95-0.85 (t, 3H). FAB Mass=454, [(M+H)⁺. Elementalanalysis: responding to the expected formula C₁₆H₃₂N₂O₄ K.F.=1.74 %water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol 7/water10.5/acetic acid 10.5) Rf=0.65. HPLC: SGE-SCX column (5 μm, 250×4 mm),mobile phase 0.05M (NH₄)H₂PO₄:CH₃CN 60:40, detector: RI, UV 205 nm,RT=9.0 min.

EXAMPLE 7 R,S-4-trimethylammonium-3-(nonyloxycarbonyl)-aminobutyrate (ST1285) (2b, FIG. 1)

Step A

The product was prepared as disclosed in Example 6, step A, using nonylchloroformate

Yield: 50%. TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol 7/water10.5/acetic acid 10.5)/acetone 7:3 Rf=0.71. HPLC: SGE-SCX column (5 μm,250×4 mm), mobile phase 50 mM (NH₄)H₂PO₄:CH₃CN 60:40, pH 4.0, detector:RI, UV 205 nm, RT=10.417 min. ¹H-NMR (300 MHz; CD₃OD): δ: 4.54-4.44 (m,1H), 4.1-4.02 (m, 2H), 3.96-3.86 (m, 2H), 3.6-3.5 (m, 2H), 3.2 (s, 9H),2.72-2.66 (m, 2H), 2-1.86 (m, 1H), 1.66-1.56 (m, 2H), 1.38-1.26 (m,14H), 0.96-0.94 (d, 6H), 0.92-0.86 (t, 3H).

Step B

The product was prepared as disclosed in Example 6, step B.

Yield 80%. M.p.=160° C. dec. ¹H-NMR (300 MHz; CD₃OD): δ: 4.5-4.35 (m,1H), 4.1-4.0 (t, 2H), 3.55-3.45 (d, 2H), 3.2 (s, 9H), 2.45-2.35 (d, 2H),1.7-1.5 (m, 2H), 1.4-1.2 (m, 14H), 0.9-0.8 (t, 3H). Elemental analysis:responding to the expected formula C₁₇H₃₄N₂O₄ K.F.=1.3 % water. TLCsilica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol 7/water 10.5/acetic acid10.5); Rf=0.62. HPLC: SGE-SCX column (5 μm, 250×4 mm), mobile phase0.05M (NH₄)H₂PO₄:CH₃CN 60:40, detector: RI, UV 205 nm, RT=7.56 mm.

The following Examples 8-9 are further illustrated by FIG. 2.

EXAMPLE 8 R,S-4-trimethylammonium-3-octyloxybutyrate (ST 1207) (6a, FIG.2)

Step A

39 g (0.3 moles) octyl alcohol were dissolved in 25 ml toluene and 14.5ml (0.107 moles) ethyl chloroacetate and 8 ml Thionyl chloride wereadded thereto at −15° C. At the end of the addition, the reactionmixture was left to stand for 4 hours at room temperature. Ethyl acetatewas then added and the solution was washed three times with 1N NaOH andsubsequently with water. The organic phase was treated with anhydroussodium sulfate, filtered and vacuum-evaporated to dryness. The productwas purified on silica gel chromatographic column, eluting with gradientfrom hexane alone to hexane/ethyl ether 95:5. The product was obtainedwith 80% yield.

TLC silica gel hexane/ethyl ether 85:15; Rf=0.75. ¹H-NMR (300 MHz;CDCl₃): δ: 4.2-4.09 (q, 2H), 3.80 (s, 2H), 3.4-3.5 (dd, 2H), 2.85 (s,2H), 1.60-1.58 (m, 2H), 1.4-1.2 (m, 10H), 0.90-0.80 (t, 3H). Elementalanalysis: responding to the expected formula C₂₂H₃₃ ClO₄.

Step B

9 ml BF₃.Et₂O were dropped to a mixture of 26.8 g (0.066 moles) of theproduct obtained in the preceding step A and 13.5 ml triethylsilane at0° C. At the end of the addition, the reaction mixture was refluxed for4 hours. After cooling, ether was added and the solution was washedtwice with NaOH 1N, then water; the organic phase was dried overanhydrous sodium sulfate, filtered and vacuum-evaporated to dryness. Anoil was obtained, which was purified on silica gel chromatographiccolumn, eluting with gradient from hexane alone to hexane/ethyl ether95:5. The product was obtained with a 70% yield.

TLC silica gel hexane/ethyl ether 90:10; Rf=0.47. ¹H-NMR (300 MHz;CDCl₃): δ: 4.2-4.09 (dd, 2H), 4.0-3.85 (m, 1H), 3.62-3.40 (m, 4H),2.70-2.50 (dd, 2H), 1.55-1.50 (m, 2H), 1.4-1.2 (m, 10H), 0.90-0.80 (t,3H). Elemental analysis: responding to the expected formula C₁₄H₂₇ClO₃

Step C

5.2 g (0.08 moles) NaN₃ and a catalytic amount of tetrabutyl ammoniumbromide were added to a solution of 11.4 g (0.041 moles) productobtained in the preceding step B. The reaction mixture was left forthree nights at 60° C. The solution was vacuum-evaporated to dryness. Athick dark solution was obtained, which was purified on silica gelchromatographic column, eluting with gradient from hexane alone tohexane/ethyl ether 95:5. The product was obtained with a 83% yield.

TLC silica gel hexane/ethyl ether 95:5; Rf=0.23. ¹H-NMR (300 MHz;CDCl₃): δ: 4.2-4.09 (dd, 2H), 4.0-3.80 (m, 1H), 3.60-3.40 (dd, 2H),3.40-3.20 (dd, 2H), 2.70-2.40 (dd, 2H), 1.60-1.40 (m, 2H), 1.4-1.1 (m,10H), 0.90-0.80 (t, 3H). Elemental analysis: responding to the expectedformula C₁₄H₂₇N₃O₃.

Step D

The product obtained in the preceding step C (15.39 g, 0.054 moles) wasdissolved in 31 ml of acetic acid and the resulting solution wassubjected to catalytic hydrogenation with 10% Pd/C at 60 psi for 7hours. The reaction progress was checked by TLC, until disappearance ofthe starting product (hexane/ethyl ether 95:5). Thereafter, formaldehydewas added (4.6 ml, 0.167 moles) followed by 10% Pd/C and the mixture washydrogenated at 30 psi for 2 days. The catalyst was filtered off and themixture was vacuum-dried. A pale yellow liquid was obtained, which wastaken up with methylene chloride, washed with IN NaOH, then water, thenNaCl saturated solution; the organic phase was dried over anhydroussodium sulfate, filtered and vacuum-evaporated to dryness. A thick oilwas obtained. The product was obtained with a 98% yield.

TLC silica gel AcOEt/MeOH/NH₃ 90:10:3; Rf=0.42. ¹H-NMR (300 MHz; CDCl₃):δ: 4.2-4.09 (dd, 2H), 3.85-3.80 (m, 1H), 3.60-3.40 (dd, 2H), 2.65-2.40(dd, 2H), 2.40-2.20 (dd, 2H), 2.20 (s, 6H), 1.60-1.40 (m, 2H), 1.4-1.1(m, 10H), 0.90-0.80 (t, 3H). Elemental analysis: responding to theexpected formula C₁₆H₃₆NO₃.

Step E

The product obtained in the preceding step D (15.21 g, 0.053 moles) wasdissolved in 98 ml THF and 8 ml methyl iodide were added thereto. Thereaction progress was left overnight at room temperature. The mixturewas vacuum-evaporated to dryness. A thick oil was obtained. The productwas obtained with a 98% yield.

TLC silica gel AcOEt/MeOH/NH₃ 90:10:3; Rf=0.10. ¹H-NMR (300 MHz; CDCl₃):δ: 4.45-4.3 (m, 1H), 4.2-4.09 (dd, 2H), 3.75-3.30 (m, 2H), 3.5 (s, 9H),2.75-2.60 (dd, 2H), 1.60-1.45 (m, 2H), 1.30-1.15 (m, 10H), 0.90-0.80 (t,3H). Elemental analysis: responding to the expected formula C₁₆H₃₉INO₃.

Step F

The product obtained in the preceding step E, was hydrolysed onAmberlyst IRA 402 resin (OH⁻ activated form) eluting with water. Waterwas evaporated to dryness; the resulting solid was treated withisopropyl alcohol three times. A white solid was obtained.

Yield=93% M.p.=106° C. dec. ¹H-NMR (300 MHz; MeOD): δ: 4.30-4.15 (m,1H), 3.70-3.60 (dd, 1H), 3.50-3.40 (m, 2H), 3.20 (s, 9H), 2.75-2.65 (dd,1H), 2.20-2.10 (dd, 1H), 1.60-1.50 (m, 2H), 1.40-1.20 (m, 10H), 0.9-0.8(t, 3H). Elemental analysis: responding to the expected formulaC₁₅H₃₁NO₃. K.F.=5.7 % water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropylalcohol 7/water 10.5/acetic acid 10.5); Rf=0.7. HPLC: SGE-SAX column (5μm, 250×4 mm), mobile phase 0.025M (NH₄)H₂PO₄:CH₃CN 30:70, detector: RI,UV 205 nm, flow=0.75 ml/min, RT=5.85 min. MS-FAB+glycerol matrix=274.

EXAMPLE 9 R,S-4-trimethylammonium-3-tetradecyloxybutyrate (ST 1228) (6b,FIG. 2)

Step A

The product was prepared as in example 8, step A using tetradecylalcohol. The product was obtained with 73% yield.

TLC silica gel hexane/ethyl ether 95:5; Rf=0.63. ¹H-NMR (300 MHz;CDCl₃): δ: 4.2-4.09 (q, 2H), 3.80 (s, 2H), 3.4-3.5 (dd, 2H), 2.85 (s,2H), 1.60-1.58 (m, 2H), 1.4-1.2 (m, 22H), 0.90-0.80 (t, 3H). Elementalanalysis: responding to the expected formula C₃₄H₆₇ClO₄.

Step B

The product was prepared as in example 8, step B. The product 2b, shownin FIG. 2, was obtained with a 72% yield.

TLC silica gel hexane/ethyl ether 95:5; Rf=0.4. ¹H-NMR (300 MHz; CDCl₃):δ: 4.2-4.09 (dd, 2H), 4.0-3.85 (m, 1H), 3.62-3.40 (m, 4H), 2.70-2.50(dd, 2H), 1.55-1.50 (m, 2H), 1.4-1.2 (m, 22H), 0.90-0.80 (t, 3H).Elemental analysis: responding to the expected formula C₂₀H₃₉O₃.

Step C

The product was prepared as in example 8, step C. The product wasobtained with 79% yield.

TLC silica gel hexane/ethyl ether 90:10; Rf=0.36. ¹H-NMR (300 MHz;CDCl₃): δ: 4.2-4.09 (dd, 2H), 4.0-3.30 (m, 1H), 3.60-3.40 (dd, 2H),3.40-3.20 (dd, 2H), 2.70-2.40 (dd, 2H), 1.60-1.40 (m, 2H), 1.4-1.1 (m,22H), 0.90-0.80 (t, 3H). Elemental analysis: responding to the expectedformula C₂₀H₃₉N₃O₃.

Step D

The product was prepared as in example 8, step D. The product wasobtained with a 98% yield.

TLC silica gel AcOEt/MeOH/NH₃ 90:10:3; Rf=0.72. ¹H-NMR (300 MHz; CDCl₃):δ: 4.2-4.09 (dd, 2H), 3.85-3.80 (m, 1H), 3.60-3.40 (dd, 2H), 2.65-2.42(dd, 2H), 2.38-2.20 (dd, 2H), 2.18 (s, 6H), 1.60-1.40 (m, 2H), 1.4-1.1(m, 22H), 0.90-0.80 (t, 3H). Elemental analysis: responding to theexpected formula C₂₂H₄₅NO₃.

Step E

The product was prepared as in example 8, step E. The product wasobtained with a 99% yield.

TLC silica gel AcOEt/MeOH/NH₃ 90:10:3; Rf=0.15. ¹H-NMR (300 MHz; CDCl₃):δ: 4.45-4.3 (m, 1H), 4.2-4.09 (dd, 2H), 3.75-3.30 (m, 2H), 3.5 (s, 9H),2.75-2.60 (dd, 2H), 1.60-1.45 (m, 2H), 1.30-1.15 (m, 22H), 0.90-0.80 (t,3H). Elemental analysis: responding to the expected formula C₂₃H₄₈INO₃.

Step F

The product was prepared as in example 8, step F. The product wasobtained with a 99% yield.

M.p.=106° C. dec. ¹H-NMR (300 MHz; DMSO-D6): δ: 4.10-4.0 (m, 1H),3.60-3.20 (m, 4H), 3.05 (s, 9H), 2.40-2.30 (dd, 1H), 1.80-1.70 (dd, 1H),1.50-1.40 (m, 2H), 1.30-1.15 (m, 22H), 0.9-0.8 (t, 3H). Elementalanalysis: responding to the expected formula C₂₁H₄₃NO₃. K.F.=6.4% water.TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol 7/water 10.5/aceticacid 10.5); Rf=0.6. HPLC: SGE-SCX column (5 μm, 250×4 mm), mobile phase0.05M (NH₄)H₂PO₄:CH₃CN 40:60, detector: RI, UV 205 nm, flow=0.75 ml/min,RT=4.38 min. MS-FAB+glycerol matrix=358.3

The following Examples 10-11 are further illustrated by FIGS. 3a-b.

EXAMPLE 10 R,S-1-guanidinium-2-tetradecyloxy-3-(tetrazolate-5-yl)propane(ST 1263) (10, FIG. 3 b)

Step A

6.65 g (0.0179 moles) of the intermediate prepared in Example 9, step Cwere dissolved in 10 ml of methanol and 10 ml of 4N NaOH were added tothe solution. The reaction was left to stand for 16 hours at roomtemperature. 20 ml 6N HCl were added to the solution, which wasextracted with ethyl acetate. The organic phase was dried over anhydroussodium sulfate, filtered and vacuum concentrated. The product wasobtained as a white solid with a 95.6% yield.

TLC silica gel hexane/ethyl ether 1:1; Rf=0.5. M.p.=42-45° C. ¹H-NMR(300 MHz; CD₃OD): δ: 3.9-3.8 (m, 1H), 3.56-3.48 (m, 2H), 3.42-3.26 (dd,2H), 2.68-2.5 (m, 2H), 1.6-1.5 (m, 2H), 1.4-1.2 (s, 22H), 0.90-0.80 (t,3H). Elemental analysis: responding to the expected formula C₁₈H₃₅N₃O₃.

Step B

At 0° C., 4.96 ml TEA were dropped into a solution containing 2.79 g(8.19 mmoles) of the compound obtained in step A, aminopropionitrile(0.58 g, 8.2 mmoles) and DEPC (diethylphosphocyanydate ) (1.71 ml) in4.2 ml of anhydrous DMF. The reaction was left to stand for 1 hour atroom temperature. The solvent was evaporated and the residue wasdissolved in ethyl acetate, washed twice with water, then with a NaClsaturated solution. The organic phase was dried over anhydrous sodiumsulfate, filtered and vacuum concentrated. The product was obtained andpurified through a silica gel column with hexane: ethyl ether(7:3/1:1/3:7).

Yield: 71%. TLC silica gel ethyl ether 100%; Rf=0.42. ¹H-NMR (300 MHz;CDCl₃): δ: 6.6-6.4 (m, 1H), 3.9-3.8 (m, 1H), 3.60-3.4 (m, 5 H), 3.3-3.2(dt, 1H), 2.7-2.6 (t, 2H), 2.6-2.4 (dd, 2H), 1.6-1.5 (m, 2H), 1.4-1.2(m, 22H), 0.90-0.80 (t, 3H). Elemental analysis: responding to theexpected formula C₂₁H₃₉N₅O₂

Step C

2.99 g (0.0114 moles) triphenylphosphine and 0.2 ml water were added toa solution containing 2.99 g (7.62 mmoles) of the compound obtained instep B. The reaction was left to stand overnight at room temperature.The solvent was evaporated off and the product was obtained and purifiedthrough a silica gel column with ethyl acetate 100%, then ethylacetate:methanol:ammonia 7:3:0.3.

Yield: 65%. TLC silica gel ethyl acetate:methanol:ammonia 7:3:0.3;Rf=0.26. ¹H-NMR (300 MHz; CD₃OD): δ: 3.78-3.7 (m, 1H), 3.58-3.48 (m,4H), 2.8-2.7 (dd, 2H), 2.7-2.6 (m, 2H),. 2.5-2.3 (dd, 2H), 1.6-1.5 (m,2H), 1.4-1.3 (m, 22H), 0.90-0.80 (t, 3H). Elemental analysis: respondingto the expected formula C₂₁H₄₁N₃O₂.

Step D

1.69 g (4.6 mmoles) of the compound obtained in step C were treated with1.2 g (5.2 mmoles) (BOC)₂O and 9.2 ml 1N NaOH for 30 minutes at roomtemperature. The reaction mixture was poured into ethyl acetate andwashed four times with 1N HCl, then water and a saturated NaCl solution.The organic phase was dried over anhydrous sodium sulfate, filtered andvacuum concentrated to dryness. The product was obtained as a whitesolid.

Yield: 100%. TLC silica gel ethyl ether 100%; Rf=0.26. M.p.=83-84° C.¹H-NMR (300 MHz; CDCl₃): δ: 7.2-7.0 (m, 1H), 4.9-4.8 (m, 1H), 3.8-3.6(m, 1H), 3.5-3.4 (dt, 4H), 3.2-3.0 (m, 2H), 2.6 (t, 2H), 2.4 (d, 2H),1.5 (m, 2H), 1.4 (s, 9H), 1.4-1.2 (m, 22H), 0.90-0.80 (t, 3H). Elementalanalysis: responding to the expected formula C₂₆H₄₉N₃O₄.

Step E

The product obtained in step D (1.19 g, 2.56 mmoles) was dissolved into12 ml of anhydrous THF, under -argon atmosphere, then 3.062 g oftriphenylphosphine, 1.54 ml of triethylsilylazido and 4.9 ml of DEAD(diethylazodicarboxylate) were dropped at 0° C. within three days, untildisappearance of the starting product. The mixture was then treated withan aqueous solution of cerium ammonium nitrate and diluted with CH₂Cl₂.The reaction was left to stand for 2 hours, the organic phase was washedwith a saturated NaCl solution, dried over anhydrous sodium sulfate andvacuum-dried. The residue was purified through a silica gel column withhexane/ethyl acetate (9:1/8:2/7:3). The product was obtained with a 66%yield.

TLC silica gel hexane/AcOEt 1:1; Rf=0.34. ¹H-NMR (300 MHz; CDCl₃): δ:4.95-4.8 (m, 1H), 4.7-4.5 (m, 2H), 3.9-3.8 (m, 1H), 3.50-3.40 (m, 1H),3.40-3.31 (m, 1H), 3.3-3.2 (m, 1H), 3.22-3.0 (dd, 2H), 3.10-3.0 (m, 3H),1.45-1.35 (m, 1H), 1.2 (m, 22H), 0.90-0.80 (t, 3H). Elemental analysis:responding to the expected formula C₂₅H₄₈N₆O₃

Step F

The product obtained in step E (0.969 g, 1.97 mmoles) was dissolved into13.09 ml anhydrous THF, then 13.1 ml of 3N HCl were added. The reactionmixture was left to stand for 2 hours, at 50° C. under stirring. Thereaction mixture -was vacuum-dried, the residue was taken up with CH₂Cl₂and treated with a 1N NaOH solution. The organic phase was separated,dried over anhydrous sodium sulfate and vacuum-dried. The product wasobtained with a 92% yield.

TLC silica gel AcOEt/MeOH/NH₃ 9:1:0.3 Rf=0.31. ¹H-NMR (300 MHz; CDCl₃):δ: 4.78-4.58 (m, 2H), 3.8-3.7 (m, 1H), 3.5-3.4 (m, 1H), 3.30-3.24 (m,1H), 3.24-3.18 (m, 4H), 3.05-3.0 (dd, 2H), 3.0-2.6 (dd, 2H), 1.4 (m,2H), 1.2 (m, 22H), 0.90-0.80 (t, 3H). Elemental analysis: responding tothe expected formula C₂₁H₄₀N₆O

Step G

The product obtained in step F (2.78 g, 7.1 mmoles) was dissolved into20 ml anhydrous MeOH, then 2.34 g iminomethanesulfonic acid (preparedwith well-known methods) were added within 3 days. The obtainedsuspension was vacuum-concentrated, then treated with 1N NaOH and leftunder stirring for 30 minutes. The solid was filtered, washed withwater, then acetone. The title product was obtained with a 45% yield.

TLC silica gel AcOEt/MeOH/NH₃ 7:3:0.3; Rf=0.22. M.p.=240° C. dec.

¹H-NMR (300 MHz; CD₃OD): δ: 3.90-3.75 (m, 1H), 3.6-3.4 (m, 2H),3.40-3.20 (m, 2H), 3.20-3.10 (dd, 1H), 2.95-2.85 (dd, 1H), 1.4 (m, 2H),1.2 (s, 22H), 0.90-0.80 (t, 3H).

Elemental analysis: responding to the expected formula C₁₉H₃₉N₇O.

HPLC: Spherisorb-C1 (5 μm, 250×4.6 mm), mobile phase 0.05 M KH₂PO₄:CH₃CN35:65, pH=3, flow 0.75 ml/min, detector: UV 205 nm, RT=5.51 min.

MS-FAB+glycerol matrix=382.

EXAMPLE 11R,S-1-trimethylammonium-2-tetradecyloxy-3-(tetrazolato-5-yl)propane (ST1287) (9, FIG. 3 b)

Steps A-F

The compounds were prepared as in steps A-F of Example 10.

Step H

2.79 g (7.14 mmoles) of the compound prepared in Example 10, step F weresuspended in 18 ml water and 1.47 ml HCOOH and 1.57 ml H₂CO were addedthereto. The reaction mixture was refluxed overnight, then was allowedto cool down and methylene chloride was added; pH was adjusted to 9 with0.5 N NaOH. The mixture was extracted three times with methylenechloride. The organic phase was washed with 0.5 N NaOH, water and driedover anhydrous sodium sulfate, filtered and vacuum concentrated. Theproduct was obtained as a solid with a 100% yield.

TLC silica gel AcOEt/MeOH/NH₃ 9:1:0.3; Rf=0.58. ¹H-NMR (300 MHz; CDCl₃):δ: 4.7-4.5 (m, 1H), 3.8-3.7 (m, 1H), 3.5-3.4 (m, 1H), 3.30-3.20 (m, 2H),3.10 (m, 3H), 2.45-2.35 (m, 2H), 2.30 (s, 6H), 1.4-1.3 (m, 2H), 1.2-1.0(m, 22H), 0.90-0.80 (t, 3H). Elemental analysis: responding to theexpected formula C₂₃H₄₄N₆O.

Step I

2.99 g (7.14 mmoles) of the compound obtained in step H were dissolvedin THF and 2.5 ml of CH₃I were added thereto. The reaction was left tostand for 3 hours at room temperature. The solvent was evaporated offand the solid residue was washed with hot ether, left overnight understirring, then filtered. The product was obtained.

Yield: 100%. TLC silica gel CHCl₃:iPrOH:MeOH:H₂O:CH₃COOH42:7:28:10.5:10.5; Rf=0.73. ¹H-NMR (300 MHz; CDCl₃): δ: 4.90-4.80 (m,2H), 4.70-4.55 (m, 1H), 4.40-4.25 (m, 1H), 3.80-3.60 (m, 2H), 3.60-3.40(m, 3H), 3.30 (s, 9H), 3.30-3.10 (m, 2H), 1.60-1.40 (m, 2H), 1.3-1.1 (m,22H), 0.9-0.8 (t, 3H). Elemental analysis: responding to the expectedformula C₂₄H₄₇IN₆O. MS-FAB+glycerol matrix=436.

Step L

The product obtained in step I (2.99 g, 5.33 mmoles) was dissolved inMeOH, then passed through IRA 402 resin in OH⁻ form, conditioned inMEOH. The title product was obtained as a solid, which was subsequentlytriturated with AcOEt.

Yield=88%. TLC silica gel CHCl₃:iPrOH:MeOH:H₂O:CH₃COOH(42:7:28:10.5:10.5)/acetone 8:2; Rf=0.73. TLC silica gelCHCl₃:iPrOH:MeOH:H₂O:CH₃COOH 42:7:28:10.5:10.5; Rf=0.73. M.p.=180° C.dec. ¹H-NMR (300 MHz; CDCl₃): δ: 4.30-4.20 (m, 1H), 3.90-3.70 (m, 2H),3.60-3.55 (m, 1H), 3.50-3.30 (m, 4H), 3.25 (m, 1H), 3.0-2.9 (m, 1H),1.60-1.40 (m, 2H), 1.3-1.1 (m, 22H), 0.9-0.8 (t, 3H). Elementalanalysis: responding to the expected formula C₂₁H₄₃N₅O. MS-FAB+glycerolmatrix=382. K.F.=1% water HPLC: Spherisorb-C1 (5 μm, 250×4.6 mm), mobilephase 0.05 M KH₂PO₄:CH₃CN 35:65, pH=3, flow 0.75 ml/min, detector: UV205 nm, RT=5.18 min. The following Examples 12-14 are furtherillustrated by FIG. 4.

EXAMPLE 12R,S-3-quinuclidinium-2-(tetradecyloxycarbonyl)-oxy-1-propanephosphonatemonobasic (ST 1260)

Step A

In anhydrous environment, −70° C., a hexane solution of 1.6 M BuLi (14ml, 0.022 moles) was dropped into a solution of dibenzyl phosphite (5.8g, 0.022 mmoles) in THF. After 15 minutes, 1.8 ml (0.022 moles) ofepibromhydrine, dissolved in 5 ml THF, were added. After the addition,etherated BF₃ (3.6 ml, 0.022 moles) was dropped very slowly. Thereaction was left for further 3 hours at −70° C. A saturated ammoniumchloride solution was added; then the temperature was left to raise toroom temperature. This solution was extracted several times with AcOEtand the gathered organic phases were treated with saturated NaHCO₃, anddried over anhydrous sodium sulfate, filtered and vacuum concentrated.An oil was obtained, which after purification on silica gelchromatography (AcOEt/Hexane 1:1); gave 1.1 g of unreacteddibenzylphosphite and 5.3 g of product of interest.

Yield=60%. TLC silica gel AcOEt/Hexane 7:3; Rf=0.54. ¹H-NMR (300 MHz;CD₃OD): δ: 7.4-7.2 (m, 10H), 5.1-4.9 (m, 4H), 4.2-4.0 (m, 1H), 3.5-3.3(dd, 2H), 2.2-2.0 (m, 2H). Elemental analysis: responding to theexpected formula C₁₇H₂₀BrO₄P. MS-FAB+glycerol matrix-399, 400, 401, 402.

Step B

2 g (5 mmoles) of the compound obtained in step A were dissolved at 10%concentration and the solution cooled down to 0° C. 1.4 ml TEA (10mmoles) and 0.62 g (5 mmoles) DMAP (dimethylaminopyridine) were droppedthereto. Immediately after, 5.2 mmoles tetradecyl chloroformate wereadded and the temperature was left to raise to room temperature. Thereaction progress was checked on TLC and worked up at the disappearanceof the starting compound. Further chloroform was added and the reactionmixture was washed with 1N HCl and water. After drying over anhydroussodium sulfate, the solvent was evaporated off and an oil was obtained,which was purified through flash-chromatography using hexane/AcOEt 7:3as eluant. The product was obtained.

Yield: 75%. TLC silica gel hexane/AcOEt 7:3; Rf=0.31. ¹H-NMR (300 MHz;CDCl₃): δ: 7.4-7.2 (m, 10H), 5.1-4.9 (m, 5H), 4.1-3.9 (m, 2H), 3.6-3.4(dd, 2H), 2.4-2.2 (m, 2H), 1.6-1.4 (m, 2H), 1.3-1.1 (m, 22H), 0.9-0.7(t, 3H). Elemental analysis: responding to the expected formulaC₃₂H₄₈BrO₆P.

Step D

The product obtained in step B (6.39 g, 10 mmoles) was dissolved in 12ml DMF, then quinuclidine was added (2.2 g, 20 mmoles) together withTBAI (tetrabutyl ammonium iodide) in catalytic amounts (1% by weightwith respect to the substrate). The reaction was carried out at atemperature of 50° C., until the starting product disappeared. At theend of reaction, the mixture was concentrated under high vacuum,obtaining a semisolid containing the product. The latter was purifiedthrough silica gel flash-chromatography, using CHCl₃/MeOH 8:3. Theproduct was obtained.

Yield=15%. TLC silica gel CHCl₃:iPrOH:MeOH:H₂O:CH₃COOH(42:7:28:10.5:10.5)/acetone 8:2; Rf=0.8. ¹H-NMR (300 MHz; MeOD): δ:7.4-7.1 (m, 50H), 5.3-5.1 (m, 1H), 4.9-4.8 (d, 2H), 4.1-4.0 (m, 2H),3.8-3.4 (m, 2H), 3.4-3.2 (m, 6H), 2.2-1.7 (m, 9H), 1.6-1.4 (m, 2H),1.3-1.1 (m, 22H), 0.9-0.7 (t, 3H). Elemental analysis: responding to theexpected formula C₃₂H₅₄NO₆P. MS-FAB+glycerol matrix=580.

Step E

The product obtained in step D was dissolved in MeOH, then 10% Pd/C (5%by weight with respect to the substrate) was added; the dispersion washydrogenated (60 psi) at room temperature for 18 hours. At the end, thedispersion was filtered through celite and concentrated to dryness. Thetitle product was obtained without further purifications.

Yield=99%. TLC silica gel CHCl₃:iPrOH:MeOH:H₂O: CH₃COOH(42:7:28:10.5:10.5)/acetone 8:2; Rf=0.57. ¹H-NMR (300 MHz; D₂O): δ:5.5-5.3 (m, 1H), 4.2-4.1 (m, 2H), 4.0-3.4 (m, 8H), 2.2-1.7 (m, 9H),1.60-1.40 (m, 2H), 1.3-1.1 (m, 22H), 0.9-0.7 (t, 3H). Elementalanalysis: responding to the expected formula C₂₅H₄₈NO₆P. MS-FAB+glycerolmatrix=490. K.F.=7% water HPLC: Spherisorb-C1 (5 μm, 250×4.6 mm), mobilephase 0.075 M KH₂PO₄:CH₃CN 60:40, flow 0.75 ml/min, detector: RI, UV 205nm, RT=16.53 min.

EXAMPLE 13R,S-3-trimethylammonium-2-(nonylaminocarbonyl)-oxy-1-propanephosphonatemonobasic (ST 1286)

Step A

The product was prepared as disclosed in step A of Example 12.

Step C

The product obtained in the previous step (4 g, 10 mmoles) was dissolvedin CH₂Cl₂ (10% solution) and etherated BF₃ (1.6 ml) and nonyl isocyanate(3.38 g, 20 mmoles) were added at room temperature. The reaction wasworked up after 30 minutes, firstly adding further CH₂Cl₂, then washingthe organic phase with 1N NaOH. several times. The product was purifiedon silica gel flash-chromatography (Hexane/AcOEt 7:3).

Yield=85%. TLC silica gel AcOEt/Hexane 6:4; Rf=0.28. ¹H-NMR (300 MHz;CDCl₃): δ: 7.4-7.2 (m, 10H), 5.1-4.9. (m, 5H), 4.6-4.2 (m, 1H), 3.7-3.5(dd, 2H), 3.2-3.0 (m, 2H), 2.4-2.2 (m, 2H), 1.5-1.3 (m, 2H), 1.3-1.1 (m,12H), 0.9-0.7 (t, 3H). Elemental analysis: responding to the expectedformula C₂₇H₄₀BrNO₅P.

Step F

The compound obtained in the preceding step (5.68 g, 10 mmoles) wasdissolved in DMF (11 ml), together with TBAI (tetrabutyl ammoniumiodide) in catalytic amounts (1% w/w with respect to the substrate).This solution was saturated with gaseous trimethylamine. The reactionwas carried out at 50° C., until the starting compound disappeared. Atthe end of the reaction, the solution was high vacuum-concentrated,obtaining a semisolid, containing the product. The latter was isolatedand purified through silica gel flash-chromatography using a gradientfrom CH₂Cl₂ only to CH₂Cl₂:MeOH 1.1. The product was obtained.

Yield: 25%. TLC silica gel CHCl₃:iPrOH:MeOH:H₂O:CH₃COOH(42:7:28:10.5:10.5)/acetone 8:2; Rf=0.73. ¹H-NMR (300 MHz; CDCl₃): δ:7.5-7.2 (m, 5H), 5.5-5.4 (m, 1H), 4.9-4.8 (m, 4H), 4.0-3.6 (m, 2H),3.2-3.1 (s, 9H), 2.2-2.1 (s, 9H), 2.0-1.8 (m,.2H), 1.5-1.4 (m, 2H),1.4-1.2 (m, 12H), 0.9-0.7 (t, 3H). Elemental analysis: responding to theexpected formula C₂₇H₄₂N₂O₅P. MS-FAB+glycerol matrix=457.

Step G

The product obtained in step F was dissolved in MeOH, then 10% Pd/C (5%by weight with respect to the substrate) was added; the dispersion washydrogenated (60 psi) at room temperature for 18 hours. At the end, thedispersion was filtered through celite and concentrated to dryness. Thetitle product was obtained without further purifications.

Yield=99%. TLC silica gel CHCl₃:iPrOH:MeOH:H₂O:CH₃COOH(42:7:28:10.5:10.5)/acetone 8:2; Rf=0.31. ¹H-NMR (300 MHz; D₂O): δ:5.6-5.5 (m, 1H), 4.1-3.5 (m, 2H), 3.2-3.1 (s, 9H), 3.1-3.0 (m, 2H),2.2-1.7 (m, 2H), 1.5-1.4 (m, 2H), 1.4-1.2 (m, 12H), 0.9-0.7 (t, 3H).Elemental analysis: responding to the expected formula C₁₅H₃₅N₂O₅P.MS-FAB+glycerol matrix=367. K.F.=3% water. HPLC: Spherisorb-C1 (5 μm,250×4.6 mm), mobile phase 0.05 M (NH₄)H₂PO₄:CH₃CN 35:65, flow 0.75ml/min, detector: RI, UV 205 nm, RT=7.31 min.

EXAMPLE 14R,S-3-pyridinium-2-(nonylaminocarbonyl)-oxy-1-propanephosphonic acidchloride (ST 1268)

Step A

The product was prepared as disclosed in step A of Example 12.

Step C

The product was prepared as disclosed in step C of Example 13.

Step H

The compound obtained in the preceding step (5.68 g, 10 mmoles) wasdissolved in anhydrous pyridine (50% solution), together with TBAI(tetrabutyl ammonium iodide) in catalytic amounts (1% w/w with respectto the substrate). The reaction was carried out at 50° C., until thestarting compound disappeared. At the end of the reaction, the solutionwas high vacuum-concentrated, obtaining a semisolid, containing theproduct, which was isolated and purified through silica gelflash-chromatography using a gradient from CH₂Cl₂ only to CH₂Cl₂:MeOHfrom 9:1 to 1:1.

Yield: 20%. TLC sica gel CHCl₃:iPrOH:MeOH:H₂O:CH₃COOH(42:7:28:10.5:10.5)/acetone 8:2; Rf=0.73. ¹H-NMR (300 MHz; CDCl₃): δ:9.4-9.3 (d, 2H), 8.2-8.1 (t, 1H), 7.9-7.8 (t, 2H), 7.3-7.1 (m, 5H),5.3-5.1 (m, 3H), 4.9-4.8 (m, 2H), 3.0-2.9 (m, 2H), 2.2-1.6 (m, 2H),1.4-1.2 (m, 2H), 1.3-1.1 (m, 12H), 0.9-0.7 (t, 3H). Elemental analysis:responding to the expected formula C₂₄H₃₈N₂O₅P. MS-FAB+glycerolmatrix=477.

Step I

The product obtained in step H (4.76 g, 10 mmoles) was dissolved in 100ml CH₂Cl₂ and 20 mmoles TMSI (trimethylsilyl iodide) were added to theresulting solution. After 30 minutes, the reaction was finished; 0.5 mlwater were added to the mixture, which was concentrated to dryness. Thefinal product was purified and isolated by RP-18 silica gelchromatography, using a gradient water/methanol 9:1 to methanol 100%.The solid was dissolved in water and passed through IRA 402 resin (Cl-activated). ST 1268 was obtained.

Yield=80%. M.p.=202-204° C. TLC silica gel CHCl₃:iPrOH:MeOH:H₂O:CH₃COOH(42:7:28:10.5:10.5)/acetone 8:2; Rf=0.48. ¹H-NMR (300 MHz; D₂O): δ:9.4-9.3 (d, 2H), 8.2-8.1 (t, 1H), 7.9-7.8 (t, 2H), 5.5-5.4 (m, 1H),5.2-4.8 (m, 2H), 3.0-2.9 (m, 2H), 2.2-2.0 (m, 2H), 1.4-1.1 (m, 14H),0.9-0.7 (t, 3H). Elemental analysis: responding to the expected formulaC₁₈H₃₂N₂ ClO₅P. MS-FAB+glycerol matrix=387. K.F.=6% water. HPLC:Spherisorb-C1 (5 μm, 250×4.6 mm), mobile phase 0.050 M KH₂PO₄:CH₃CN35:65, flow 0.75 ml/min, detector: RI, UV 205 nm, RT=5.61 min.

EXAMPLE 15 R-4-trimethylammonium-3-(tetradecylcarbamoyl)-amino Butyrate(ST 1326)

The product was prepared as disclosed in Example 1, starting fromtetradecyl isocyanate and R-aminocarnitine, inner salt, except the crudeproduct was obtained by precipitation with ethyl ether, from thereaction mixture, directly washed with ethyl ether and purified on asilica gel chromatographic column.

Yield 57%. M.p.: 160-162° C. [α]₂₀ ^(D)=−21.1° (c=0.5, MeOH). ¹H-NMR(300 MHz; CD₃OD): δ: 4.52 1H), 3.60 (dd, 1H), 3.48 (d, 1H), 3.20 (s,9H), 3.10 (t, 2H), 2.40 (m, 2H), 1.45 (m, 2H), 1.28 (brs, 22H), 0.8(brt, 3H). ESI Mass=400, [(M+H)⁺. Elemental analysis: responding to theexpected formula C₂₂H₄₅N₃O₃. K.F.=2.5% water. TLC silica gelCHCl₃:iPrOH:MeOH:H₂O:CH₃COOH 42:7:28:10.5: 10.5; Rf=0.50. HPLC: SGE-SCXcolumn (5 μm, 250×4 mm), T=30° C., mobile phase 0.05 M (NH₄)H₂PO₄:CH₃CN75:25, pH=4.9 (as such), flow 0.75 ml/min, detector: RI, UV 205 nm,RT=13.63 min.

EXAMPLE 16 R-4-trimethylammonium-3-(undecylcarbamoyl)-aminobutyrate (ST1327)

The product was prepared as disclosed in Example 1, starting fromundecyl isocyanate and R-aminocarnitine, inner salt, purified on asilica gel chromatographic column and further purified by precipitationfrom acetonitrile.

Yield 50%. M.p.: 149-150.2° C. [α]₂₀ ^(D)=−21.16° (c=1, MeOH). ¹H-NMR(300 MHz; CD₃OD): δ: 4.52 1H), 3.60 (dd, 1H), 3.48 (d, 1H), 3.20 (s,9H), 3.10 (t, 2H), 2.40 (m, 2H), 1.45 (m, 2H), 1.28 (brs, 16H), 0.8(brt, 3H). ESI Mass=358, [(M+H)⁺; Elemental analysis: responding to theexpected formula C₁₉H₃₉N₃O₃. K.F.=2.3% water. TLC silica gelCHCl₃:iPrOH:MeOH:H₂O:CH₃COOH 42:7:28:10.5: 10.5. Rf=0.50. HPLC: SGE-SCXcolumn (5 μm, 250×4 mm), T=30° C., mobile phase 0.05 M (NH₄)H₂PO₄:CH₃CN80:20, pH=4.9 (as such), flow 0.75 ml/min, detector: RI, UV 205 nm,RT=17.37 min.

EXAMPLE 17 R-4-trimethylammonium-3-(heptylcarbamoyl)-aminobutyrate (ST1328)

The product was prepared as disclosed in Example 1, starting from heptylisocyanate and R-aminocarnitine, inner salt,.purified on a silica gelchromatographic column and further purified by precipitation fromacetonitrile.

Yield 47%. M.p.: 149-150° C. [α]₂₀ ^(D)=−34.0° (c=0.97, MeOH). ¹H-NMR(300 MHz; CD₃OD): δ: 4.52 (m, 1H), 3.60 (dd, 1H), 3.48 (d, 1H), 3.20 (s,9H), 3.10 (t, 2H), 2.40 (m, 2H), 1.45 (m, 2H), 1.30 (brs, 8H), 0.8 (brt,3H). ESI Mass=302, [(M+H)⁺; Elemental analysis: responding to theexpected formula C₁₅H₃₁N₃O₃K.F.=6.17% water TLC silica gelCHCl₃:iPrOH:MeOH:H₂O:CH₃COOH 42:7:28:10.5:10.5. Rf=0.50. HPLC: SGE-SCXcolumn (5 μ, 250×4 mm), T=30° C., mobile phase 0.05 M (NH₄)H₂PO₄:CH₃CN85:15, pH=6 (H₃PO₄), flow 0.75 mI/min, detector: RI, UV 205 nm, RT=7.16min.

EXAMPLE 18 R,S-4-trimethylammonium-3-(nonylthiocarbamoyl)-aminobutyrate(ST 1329)

The product was prepared as disclosed in Example 1, starting from nonylisothiocyanate and R,S-aminocarnitine, inner salt. Chromatography wascarried out with a CHCl₃/MeOH gradient from 8:2 to 2:8.

Yield 53% M.p.: 104-107° C. ¹H-NMR (200 MHz; CD₃OD): δ: 5.45 (brm, 1H),3.75 (dd, 1H), 3.55 (d, 1H), 3.45 (brm, (2H), 3.22 (s, 9H), 2.48 (m,2H), 1.55 (m, 2H), 1.30 (brs,-12H), 0.90 (brt, 3H). ESI Mass=346,[(M+H)⁺; Elemental analysis: responding to the expected formulaC₁₇H₃₅N₃O₂S K.F.=2.6% water; TLC silica gel CHCl₃:iPrOH:MeOH:H₂O:CH₃COOH42:7:28:10.5:10.5. Rf=0.74; HPLC: SGE-SCX column (5 μm, 250×4 mm), T=30°C., mobile phase 0.05 M (NH₄)H₂PO₄:CH₃CN 85:15, pH=6.0 (H₃PO₄), flow0.75 ml/min, detector: RI, UV 205 nm, RT=8.87 min.

EXAMPLE 19 R-4-trimethylammonium-3-(nonylcarbamoyl)-aminobutyrate (ST1283)

The product was prepared as disclosed in Example 1, starting from nonylisocyanate and R-aminocarnitine, inner salt. M.p.: 146-147° C. [α]₂₀^(D)=−13.4° (c=0.5, H₂O). Elemental analysis: responding to the expectedformula C₁₇H₃₅N₃O₃ K.F.=2.8% water. Remaining physico-chemical data werecoincident with those of racemic ST1251 (Example 1).

EXAMPLE 20 S-4-trimethylammonium-3-(nonylcarbamoyl)-aminobutyrate (ST1338)

The product was prepared as disclosed in Example 1, starting from nonylisocyanate and S-aminocarnitine, inner salt.

M.p.: 146-147° C. [α]₂₀ ^(D)=+16.7° (c=0.43, H₂O). ¹H-NMR (300 MHz;CD₃OD): δ: 4.52 (m, 1H), 3.60 (dd, 1H), 3.45 (d, 1H), 3.18 (s, 9H), 3.10(t, 2H), 2.40 (m, 2H), 1.45 (m, 2H), 1.28 (brs, 12H), 0.90 (brt, 3H).ESI Mass=330, [(M+H)⁺; Elemental analysis: responding to the expectedformula C₁₇H₃₅N₃O₃ K.F.=1.8% water. Remaining physico-chemical data werecoincident with those of racemic ST1251 (Example 1).

EXAMPLE 21 S-4-trimethylammonium-3-(tetradecylcarbamoyl)-aminobutyrate(ST 1340)

The product was prepared as disclosed in Example 1, starting fromtetradecyl isocyanate and S-aminocarnitine, inner salt, except the crudeproduct was obtained by precipitation with ethyl ether, from thereaction mixture, directly washed with ethyl ether and purified on asilica gel chromatographic column.

Yield=57%; M.p.: 166-167° C. [α]₂₀ ^(D)=+20.7° (c=0.5, MeOH). Elementalanalysis: responding to the expected formula C₂₂H₄₅N₃O₃ K.F. =1.7%water. Remaining physico-chemical data were coincident with those ofracemic ST1326 (Example 15).

EXAMPLE 22 IsobutylR,S-4-trimethylammonium-3-tetradecylamino-aminobutyrate (ST 1252)

R,S-4-trimethylammonium-3-tetradecylamino-aminobutvrate is isobutylEster Acetate

Isobutyl ester of racemic aminocarnitine (5 g, 0.0198 moles) andtetradecanal (4.6 g, 0.0217 moles) were dissolved into 250 ml methanol.Glacial acetic acid (1.13 ml, 0.198 moles) and 1 g 10% Pd/C were added.The mixture was hydrogenated at 30 psi overnight. After filtration oncelite, the solution was vacuum-concentrated. A pale yellow oil wasobtained, which was purified through a silica gel column, elutingfirstly with AcOEt, then AcOEt/MeOH 9:1. 4 g of product were obtained.

Yield=47%;

TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol 7/water 10.5/aceticacid 10.5)/methyl acetate 7:3

Rf=0.74. ¹H-NMR (300 MHz; CD₃OD): δ: 3.92-3.90 (d, 2H), 3.64-3.58 (m,1H), 3.50-3.30 (m, 2H), 2.80-2.50 (m, 4H), 2.0-1.9 (m, 1H), 2.6-2.4 (m,2H), 1.3 (s, 22H), 0.98-0.82 (m, 9H).

R,S-4-trimethylammonium-3-tetradecylamino-aminobutyrate

The isobutyl ester ofR,S-4-trimethylammonium-3-tetradecylamino-aminobutyric acid, acetatesalt, (3.3 g) was hydrolysed on Amberlyst IRA 402 resin (OH⁻ activatedform) and eluted with water. Water was evaporated to dryness underreduced pressure; the resulting white solid was washed with methanol,filtered and vacuum-dried. 1.95 g of product were obtained.

Yield 70% M.p.=160° C. dec. ¹H-NMR (300 MHz; CD₃OD): δ: 4.4 (m, 1H),3.40-3.35 (m, 3H), 3.2 (s, 9H), 2.80-2.72 (m, 1H), 2.56-2.42 (m, 2H),2.27-2.16 (m, 1H), 1.55-1.40 (m, 2H), 1.3 (s, 22H), 0.92-0.85 (t, 3H).Elemental analysis: responding to the expected formula C₂₁H₄₄N₂O₂K.F.=1.93 % water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol7/water 10.5/acetic acid 10.5) Rf=0.5. HPLC: SGE-SCX column (5 μm, 250×4mm), mobile phase 0.05M (NH₄)H₂PO₄:CH₃CN 60:40, pH=4, flow=0.75 ml/min;detector: RI, UV 205 nm, RT=30.017 min.

EXAMPLE 23 R,S-4-trimethylammonium-3-octylaminobutyrate (ST 1254)

R,S-4-trimethylammonium-3-octylamino-aminobutyrate Isobutyl EsterAcetate

Isobutyl ester of racemic aminocarnitine chloride, (5 g, 0.0198 moles)and octanaldehyde (2.79 g, 0.0217 moles) were dissolved into 250 mlmethanol. Glacial acetic acid (1.13 ml, 0.198 moles) and 1 g 10% Pd/Cwere added. The mixture was hydrogenated at 30 psi overnight. Afterfiltration on celite, the solution was vacuum-concentrated. 8.5 gproduct were obtained, subsequently purified through a silica gelcolumn, eluting firstly with AcOEt, then AcOEt/MeOH (9:1; 8.5:1.5). 3 gof product were obtained.

Yield=40%; TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol 7/water10.5/acetic acid 10.5) Rf=0.54. ¹H-NMR (300 MHz; CD₃OD): δ: 3.92-3.90(d, 2H), 3.64-3.58 (m, 1H), 3.50-3.30 (m, 2H), 2.80-2.50 (m, 4H),2.0-1.9 (m, 1H), 2.6-2.4 (m, 2H), 1.3 (s, 10H), 0.98-0.82 (m, 9H).

R,S-4-trimethylammonium-3- octylaminobutyrate

The isobutyl ester ofR,S-4-trimethylammonium-3-tetradecylamino-aminobutyric acid, acetatesalt, (2.8 g, 0.00719) was hydrolysed on Amberlyst IRA 402 resin (OH⁻activated form) and eluted with water. Water was evaporated to drynessunder reduced pressure; the resulting white solid was washed withmethanol, filtered and vacuum-dried. 1.8 g of product were obtained.

Yield 70% M.p.=140° C. dec. ¹H-NMR (300 MHz; CD₃OD): δ: 3.42-3.30 (m,3H), 3.2 (s, 9H), 2.85-2.70 (m, 1H), 2.60-2.40 (m, 2H), 2.30-2.20 (m,1H), 1.55-1.40 (m, 2H), 1.3 (s, 10H), 0.92-0.85 (t, 3H). Elementalanalysis: responding to the expected formula C₁₅H₃₂N₂O₂ K.F.=2.8 %water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol 7/water10.5/acetic acid 10.5) Rf=0.32. HPLC: SGE-SCX column (5 μm, 250×4 mm),mobile phase 0.05M (NH₄)H₂PO₄: CH₃CN 40:60, pH=4, flow=0.75 ml/min;detector: RI, UV 205 nm, RT=43.20 min.

EXAMPLE 24 R,S-4-trimethylammonium-3-(decansulfonyl)aminobutyrate (ST1364)

Aminocarnitine isobutyl ester chloride hydrochloride

Isobutyl ester of aminocarnitine, inner salt (3 g, 18.72 mmoles), wasdissolved in isobutanol (120 ml) and ice-bath cooled. Gaseous HCl wasbubbled into the solution until complete saturation and clearing of themixture. The solution was refluxed (bath temperature 130° C.) overnight.The solvent was vacuum-evaporated and the residue was triturated withEt₂O. 5.1 g of white solid were obtained.

Yield=95%; ¹H-NMR (200 MHz; D₂O): δ: 4.3 (m, 1H), 4.0 (d, 2H), 3.8 (d,2H), 3.2 (s, 9H), 3.1 (m, 2H), 2.0 (m, 1H), 0.9 (d, 6H). Elementalanalysis: responding to the expected formula C₁₁H₂₆C₂N₂O₂. K.F.=1 %water.

R,S-4-trimethylammonium-3-(decansulfonyl)-aminobutyrate

The isobutyl ester of R,S-aminocarnitine chloride, hydrochloride (1 g,3.46 mmoles) in anhydrous dichloromethane (5 ml) was added withtriethylamine (2.65 ml, 19mmoles) and decansulfonyl chloride (2.1 g,8.65 mmoles) -suspended in 3 ml anhydrous dichloromethane, at 0° C. Themixture was left under stirring for 3 days at room temperature. Thesolvent was evaporated to dryness, the residue was taken up with ethylacetate and the white precipitate of triethylamine hydrochloride wasseparated by from the solution by vacuum-filtration. The ethyl acetatesolution was vacuum-dried to give 2.8 g of a yellow oil. 71 ml 1N NaOHwere added to hydrolize the isobutyl ester, leaving the suspension understirring overnight at room temperature. The suspension was evaporatedand vacuum-dried, and the solid residue was completely dried underoil-vacuum, taken up with methanol and purified through silica gelchromatographic column, using methanol as eluant. 555 mg of product wereobtained.

Yield 44% M.p.=158° C. dec. ¹H-NMR (300 MHz; CD₃OD): δ: 4.3 (m, 1H),3.45 (m, 2H), 3.25 (s, 9H), 3.15 (m, 2H), 2.45 (d, 2H), 1.8 (m, 2H),1.45 (m, 2H), 1.4 (brs, 12H), 0.9 (brt, 3H). Elemental analysis:responding to the expected formula C₁₇H₃₆N₂O₄S Mass ESI=365 [(M+H)⁺],387[(M+Na)⁺] K.F.=3% water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropylalcohol 7/water 10.5/acetic acid 10.5) Rf=0.62. HPLC: Spherisorb-C1column (5 μm, 250×4.6 mm), mobile phase 0.05M K₂H₂PO₄:CH₃CN 35:65, pH assuch, flow=0.73 ml/min; temperature=30° C., detector: RI, UV 205 nm,RT=7.0 min.

EXAMPLE 25 R,S-4-trimethylammonium-3-(nonylsulfamoyl) aminobutyrate (ST1362)

The isobutyl ester of R,S-aminocarnitine chloride, hydrochloride (2 g,6.9 mmoles) in anhydrous dichloromethane (40 ml) was added withtriethylamine (3.8 ml, 27.6 mmoles) and dropped with SO₂Cl₂ indichloromethane (1.7 ml in 10 ml final solution) at 0° C. The mixturewas left under stirring for 3 days at room temperature, triethylamine(1.9 ml, 13.8 mmoles) and nonylamine (2.5 ml, 13.8 mmoles) were addedand the reaction mixture was left under stirring overnight at roomtemperature. The solvent was vacuum-evaporated, the residue was taken upwith ethyl acetate (100 ml) and the precipitate of triethylaminehydrochloride was separated from the solution by vacuum-filtration. Theethyl acetate solution was vacuum-dried to give 4.8 g of a yellow oil,to which were added 105 ml 1N NaOH to hydrolize the isobutyl ester. Themixture was left under stirring overnight at room temperature andvacuum-dried. The residue was completely dried under oil-vacuum. Theyellow semisolid was crystallized from chloroform. 1.26 g of productwere obtained.

Yield 50% M.p.=152° C. dec. ¹H-NMR (300 MHz; CD₃OD): δ: 4.1 (m, 1H),3.48 (d, 2H), 3.25 (s, 9H), 2.95 (m, 2H), 2.5 (t, 2H), 1.55 (t, 2H),1.45 (brs, 12H), 0.9 (brt, 3H). Elemental analysis: responding to theexpected formula C₁₆H₃₅N₃O₄S Mass ESI=366 [(M+H)⁺], 388[(M+Na)⁺]K.F.=5.8% water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol7/water 10.5/acetic acid 10.5) Rf=0.34. HPLC: Spherisorb-C1 column (5μm, 250×4.6 mm), mobile phase 0.05M KH₂PO₄:CH₃CN 35:65, pH as such,flow=0.75 ml/min; temperature=30° C., detector: RI, UV 205 nm, RT=6.68min.

EXAMPLE 26 S-4-trim ethylammonium-3-(dodecansulfonyl)aminobutyrate (ST1391)

The product was prepared as disclosed in Example 24, starting fromisobutyl ester of S-arninocarnitine chloride, hydrochloride anddodecansulfonyl chloride, to give 600 mg of product.

Yield 44% M.p.=156° C. dec. [α]_(D) ²⁰=+6° (c=0.245%, H₂O) ¹H-NMR (300MHz; CD₃OD): δ: 4.3 (m, 1H), 3.45 (m, 2H), 3.25 (s, 9H), 3.15 (m, 2H),2.45 (d, 2H), 1.8 (m, 2H), 1.45 (m, 2H), 1.4 (brs, 16H), 0.9 (brt, 3H).Elemental analysis: responding to the expected formula C₁₉H₄₀N₂O₄SK.F.=8.6% water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol7/water 10.5/acetic acid 10.5) Rf=0.65. HPLC: Spherisorb-C1 column (5 1μm, 250×4.6 mm), mobile phase 0.05M KH₂PO₄:CH₃CN 40:60, pH as such,flow=0.75 ml/min; temperature=30° C., detector: RI, UV 205 nm, RT=8.5min.

EXAMPLE 27 R-4-trimethylammonium-3-(dodecansulfonyl)aminobutyrate (ST1420)

The product was prepared as disclosed in Example 24, starting fromisobutyl ester of R-aminocarnitine chloride, hydrochloride anddodecansulfonyl chloride, to give 450 mg of product.

Yield 34% M.p.=158° C. dec. [α]_(D) ²⁰=−7° (c=0.265%, H₂O) ¹H-NMR (300MHz; CD₃OD): δ: 4.3 (m, 1H), 3.45 (m, 2H), 3.28 (s, 9H), 3.15 (m, 2H),2.45 (d, 2H), 1.8. (m, 2H), 1.45 (m, 2H), 1.3 (brs, 16H), 0.9 (brt, 3H).Elemental analysis: responding to the expected formula C₁₉H₄₀N₂O₄SK.F.=6.9% water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol7/water 10.5/acetic acid 10.5) Rf=0.66. HPLC: Spherisorb-C1 column (5μm, 250×4.6 mm), mobile phase 0.05M KH₂PO₄:CH₃CN 40:60, pH as such,flow=0.75 ml/min; temperature=30° C., detector: RI, UV 205 nm, RT=8.11min.

EXAMPLE 28 S-4-trimethylammonium-3-(undecylsulfamoyl)aminobutyrate (ST1427)

The product was prepared as disclosed in Example 25, starting fromisobutyl ester of S-aminocarnitine chloride, hydrochloride and undecylamine, except the crude product was purified on a silica gelchromatographic column, using a gradient CHCl₃: MeOH 9:1 to 1:9. Theproduct was further purified on a silica gel chromatographic column,using MeOH. 0.7 g of pure product were obtained.

Yield 38% M.p.=153° C. dec. [α]_(D) ²⁰=+4° (c=0.25%, H₂O, pH=2) ¹H-NMR(300 MHz; CD₃OD): δ: 4.1 (m, 1H), 3.48 (d, 2H), 3.25 (s, 9H), 2.95 (m,2H), 2.5 (m, 2H), 1.55 (brt, 2H), 1.45 (brs, 16H), 0.9 (brt, 3H).Elemental analysis: responding to the expected formula C₁₈H₃₉N₃O₄SK.F.=2.9% water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol7/water 10.5/acetic acid 10.5) Rf=0.68. HPLC: Spherisorb-C1 column (5μm, 250×4.6 mm), mobile phase 0.05M KH₂PO₄:CH₃CN 60:40, pH as such,flow=0.7 ml/min; temperature=30° C., detector: RI, UV 205 nm, RT=8.384min.

EXAMPLE 29 R-4-trimethylammonium-3-(undecylsulfamoyl)aminobutyrate (ST1428)

The product was prepared as disclosed in Example 25, starting fromisobutyl ester of S-aminocarnitine chloride, hydrochloride and undecylamine, except the crude product was purified on a silica gelchromatographic column, using a gradient CHCl₃: MeOH 9:1 to 1:9. Theproduct was further purified on a silica gel chromatographic column,using MeOH. 0.5 g of product were obtained.

Yield 32% M.p.=158° C. dec. [α]_(D) ²⁰=−4° (c=0.25%, H₂O, pH=2) ¹H-NMR(300 MHz; CD₃OD): δ: 4.1 (m, 1H), 3.48 (d, 2H), 3.25 (s, 9 H), 2.95 (m,2H), 2.5 (m, 2H), 1.55 (brm, 2H), 1.45 (brs, 16H), 0.9 (brt, 3H).Elemental analysis: responding to the expected formula C₁₈H₃₉N₃O₄SK.F.=4.77% water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol7/water 10.5/acetic acid 10.5) Rf=0.68. HPLC: Spherisorb-C1 column (5μm, 250×4.6 mm), mobile phase 0.05M KH₂PO₄:CH₃CN 60:40, pH as such,flow=0.7 ml/min; temperature=30° C., detector: RI, UV 205 nm, RT=8.379min.

EXAMPLE 30 R-4-trimethylammonium-3-(dodecylcarbamoyl)aminobutyrate (ST1375)

The product was prepared as disclosed in Example 1, starting fromR-aminocarnitine inner salt and dodecylisocyanate. The crude productobtained after washing with diethyl ether was purified on a silica gelchromatographic column to give 4.8 g of product.

Yield 55% M.p.=147° C. dec. [α]_(D) ²⁰=−24.6° (c=0.48%, MeOH) ¹H-NMR(300 MHz; CD₃OD): δ: 4.51 (m, 1H), 3.60 (dd, 1H), 3.45 (dd, 1H), 3.2 (s,9H), 3.1 (t, 2H), 2.4 (m, 2H), 1.45 (m, 2H), 1.3 (brs, 18H), 0.9 (t,3H). Elemental analysis: responding to the expected formula C₂₀H₄₁N₃O₃K.F.=5.4% water. TLC silica gel (CHCl₃ 42/MeOH 28/isopropyl alcohol7/water 10.5/acetic acid 10.5) Rf=0.6. HPLC: Spherisorb-C1 column (5 μm,250×4.6 mm), mobile phase 0.05M KH₂PO₄:CH₃CN 65:35, pH=5.6, flow=0.75ml/min; temperature=30° C., detector: RI, UV 205 nm, RT=8.5 min.

EXAMPLE 31R-4-trimethylammonium-3-(10-phenoxydecylcarbamoyl)aminobutyrate (ST1449)

10-Phenoxydecyl Isocyanate

A solution of 11-phenoxyundecanoyl chloride (31.1 g, 104.8 mmoles) inacetone (30 ml) was dropped into a solution of sodium azide (9.53 g,146.6 mmoles) in water (30 ml), cooled in an ice bath, keeping thesolution temperature between 10 and 15° C. After one hour, the solutionwas transferred in a separatory funnel and the lower phase (the aqueousone) was eliminated. The higher phase was transferred into a flaskcontaining 100 ml of toluene, previously warmed at 65° C. After 1.5hours, the solution was evaporated to dryness, giving 13.37 g of crudeproduct, which could be used as such in the subsequent reaction.

¹H-NMR (300 MHz; CDCl₃): δ: 7.2 (m, 2H), 6.9 (m, 3H), 3.9 (t, 2H), 3.6(t, 2H), 1.4 (m, 2H), 1.3 (m, 10H).

R-4- trimethylammonium-3-(10 -phenoxydecylcarbamoyl)-amino Butyrate

10-phenoxydecylisocyanate (25.0 g, 91.12 mmoles) was added to a solutionof aminocarnitine, inner salt (7.3 g, 45.56 mmoles) in anhydrous DMSO(350 ml) and the solution was left to stand for 60 hours at 40° C. Theresulting mixture was transferred in a 3 l Erlenmeyer flask containingethyl ether (2.5 l) and the solvent was separated by decantation of theformed precipitate, which was then taken with few chloroform,transferred into a flask and precipitated again with ethyl ether. The soobtained crude product was washed several times with ethyl ether andpurified on a silica gel chromatographic column, using a gradient CHCl₃:MeOH 9:1 to CHCl₃: MeOH 3:7 gradient until elution of impurities withhigher Rf, then eluting the product of interest with MeOH only. 13.5 gof pure product were obtained.

Yield 68% ¹H-NMR (300 MHz; CD₃OD): δ: 7.2 (m, 2H), 6.9 (m, 3H), 4.5 (m,1H), 3.9 (t, 2H), 3.6 (dd, 1H), 3.4 (dd, 1H), 3.2 (s, 9H), 3.1 (t, 2H),2.4 (m, 2H), 1.8 (m, 2H), 1.6 (m, 2H), 1.4 (m, 2H), 1.3 (m, 10H). FABMass=436, [(M+H)⁺; Elemental analysis: responding to the expectedformula C₂₄H₄₁N₃O₄ K.F.=2.3% water.

EXAMPLE 32R-4-trimethylammonium-3-(trans-β-styrenesulfonyl)aminobutyrate (ST 1448)

R-aminocarnitine isobutyl ester chloride hydrochloride

R-aminocarnitine inner salt (3 g, 18.72 mmoles) was dissolved inisobutanol (120 ml) and ice-bath cooled. Gaseous HCl was bubbled intothe solution until complete saturation and clearing of the mixture. Thesolution was refluxed (bath temperature 130° C.) overnight. The solventwas vacuum-evaporated and the residue was triturated with Et₂O. 5.1 g ofwhite solid were obtained.

Yield=95%; ¹H-NMR (200 MHz; D₂O): δ: 4.3 (m, 1H), 4.0 (d, 2H), 3.8 (d,2H), 3.2 (s, 9H), 3.1 (m, 2H), 2.0 (m, 1H), 0.9 (d, 6H). Elementalanalysis: responding to the expected formula C₁₁H₂₆Cl₂N₂O₂. K.F.=1%water.

R-4-trimethylammonium-3-(trans-β-styrenesulfonyl) -aminobutyrate

The isobutyl ester of R-aminocarnitine chloride, hydrochloride (1 g,3.46 mmoles) in anhydrous dichloromethane (5 ml) was added withtriethylamine (2.65 ml, 19 mmoles) and trans-β-styrenesulfonyl chloride(1.753 g, 8.65 mmoles) suspended in 3 ml anhydrous dichloromethane, at0° C. The mixture was left under stirring for 3 days at roomtemperature. The solvent was evaporated to dryness, the residue wastaken up with ethyl acetate (100 ml) and the white precipitate oftriethylamine hydrochloride was separated by from the solution byvacuum-filtration. The ethyl acetate solution was vacuum-dried, then 71ml 1N NaOH were added to hydrolize the isobutyl ester, leaving thesuspension under stirring overnight at room temperature. The suspensionwas evaporated and vacuum-dried, and the solid residue was completelydried under oil-vacuum, taken up with methanol and purified throughsilica gel chromatographic column, using methanol as eluant. 565 mg ofproduct were obtained.

Yield 50% ¹H-NMR (300 MHz; CD₃OD): δ: 7.8 (d, 1H), 7.5 (m, 5H), 7.3 (d,1H), 4.3 (m, 1H), 3.4 (m, 2H), 3.2 (s, 9H), 2.4 (d, 2H). Elementalanalysis: responding to the expected formula C₁₅H₂₂N₂O₄S ESI Mass=327[(M+H)⁺]

PHARMACOLOGICAL ACTIVITY Determination of CPT Inhibiting Activity.

CPT inhibition was evaluated essentially as described in Kerner, J. &Bieber, L. L. (1990) Biochemistry 29: 4326-34 on fresh mitochondrialpreparations obtained from normally fed Fischer rat liver or heart.Mitochondria were isolated from liver or heart and suspended in 75 mMsaccharose buffer, 1 mM EGTA, pH 7.5. 100 μl mitochondrial suspension,containing 50 μM [¹⁴C] palmitoyl-CoA (specific activity 10,000 DPM/mole)and 10 mM L-carnitine, were incubated at 37° C., in the presence ofscalar concentrations of the test product (0-3 mM). Reaction time: 1minute.

Table 1 shows the IC₅₀ determined.

The compounds of the present invention have higher inhibiting activitythan the one of the reference compound SDZ-CPI-975, Example 1, disclosedin EP 0 574 355.

TABLE 1 IC₅₀ of inhibition CPT1 curve in rat liver mitochondria IC₅₀Compound (μM/I) SDZ-CPI-975 17.4 ST1326 0.75 ST1327 3.2

Determination of oleate-stimulated β-hydroxybutyrate production

β-hydroxybutyrate production is an index of CPT activity. In fact, theproduction of ketone bodies, final products of mitochondrialβ-oxidation, is related to CPT activity.

Mithocondrial preparations, obtained according to the method byVenerando et al. (Am. J. Physiol. 266:C455-C461, 1994), were used.Hepatocytes are incubated at 37° C. in KRB bicarbonate buffer at pH 7.4,6 mM glucose, 1% BSA in O₂/CO₂ 95/5 atmosphere at 2.5×10⁶ cells/ml.After 40 min incubation with the test compound at differentconcentrations, the first set of samples was taken (T_(0 min)) andoleate was added (1 mM final in KRB+BSA 1.4%). After 20 minutes, thesecond withdrawal was made (T_(20 min)).

Table 2 shows the results. The data are the mean of three differentexperiments, twice carried out.

The compounds of the present invention have higher β-hydroxybutyrateinhibiting activity than the one of the reference compound SDZ-CPI-975,Example 1, disclosed in EP 0 574 355.

TABLE 2 IC₅₀ of inhibition CPT1 curve of β-hydroxybutyrate production inrat hepatocytes IC₅₀ Compound (μM/I) SDZ-CPI-975 3.7 ST1251 0.5 ST12530.9 ST1285 1.9

Glucose and β-hydroxybutyrate in Serum Fasted Rats Treated With CPTInhibitors

Normally fed Fischer rats were starved for 24 hours and subsequentlytreated with the test compounds. One hour after the treatment, theanimals were sacrificed and serum concentrations of glucose andβ-hydroxybutyrate were determined.

Table 3 shows the results. For the compound ST1326 were used doses of14.5 mg/2 ml/kg, for other test compounds, the doses are equivalent toST1326 one.

TABLE 3 β-hydroxybutyrate and glucose serum concentration in 24hours-starved rats, after one hour from intraperitoneal treatment. SDZcontrol CPI-975 ST1251 ST1253 ST1326 ST1327 ST1328 β- OHB Mean 1867119.9 99.8 118.8 133.1 93.0 169.2 s.e. 240 12.8 8.3 20.4 12.4 8.7 26.7p< — 0.001 0.001 0.001 0.001 0.001 0.001 Glu Mean 108.8 87.6 76.9 88.284.2 84.9 79.5 s.e. 6.7 1.0 2.3 3.9 2.4 1.6 1.6 p< — 0.05 0.01 0.05 0.050.05 0.05

Glucose and Insulin Levels in Diabetic Animals Treated with CPTInhibitors

C57BL/6J male rats, 5-weeks old, were provided by Ch. River. After 10days of acclimatisation in standard conditions (22±2° C.; 55±15%humidity; 15-20/h air changes; 12 hours light-dark cycle, with 700-1900lux) and with standard diet with 4RF21 feedstock (Mucedola), glycaemiawas controlled in post-absorption state (starving from 8.30 a.m. to 4.30p.m.). Blood withdrawal was carried out cutting the tail end. Glucosewas analysed in blood acid supernatant (HCLO4 0,375 N) with autoanalyzerCobas Mira S with Glucose GDH Kit (Roche).

The animals were divided in two groups, 26 mice each and fed with ahigh-fat and a low-fat diet, respectively.

After 2 months from the start of the diet, glycaemia was tested,according to the starting method. After about 3 months from the start ofthe diet, glycaemia was tested, according to the starting method andplasma insulin levels were also determined (with blood withdrawal fromend tail cutting) using [125I] Rat Insulin Kit (Amersham).

One 10 mice group fed with low-fat diet and two 10-mice groups fed withhigh -fat diet were selected One of the two high fat diet wasadministered with ST 1327 at the dose of 45 mg/Kg in deionised H2O(p.o., twice a day, 8.30 a.m. and 5.30 p.m.).administration volume was10 ml/Kg. the two remaining groups was treated with vehicle only.High-fat or low-fat diets were continued during the treatment.

After 20 days of treatment, glycaemia and plasma insulin were measured.After 43 days of treatment, the animals were sacrificed by decapitationin post-absorption state (fasting 8.30 a.m.−4.30 p.m.), 8 hours afterthe last treatment. Blood was withdrawn and serum was separated bycentrifugation and stored at −80° C. Liver, heart and skeletal muscle(upper limbs) were also extracted, frozen in dry ice-acetone and kept at−80° C.

High-fat diet determined an increase of body weight, glycaemia andinsulin, with respect to low-fat diet.

After 20 days of treatment with ST 1327, glucose and insulin levelssignificantly decreased.

Table 4 shows the results.

TABLE 4 Glucose and insulin levels in rats fed with fat-rich diet. HighFat diet High Fat diet Low fat diet Compound Control Treated ControlGlucose 248.5 ± 11.03 181.4 ± 9.63*  207.3 ± 6.84** mg/dl (10) (9) (9)Insulin 1.632 ± 0.246 0.621 ± 0.117** 0.549 ± 0.050* ng/ml (10) (9) (9)

Student's t test, * and ** indicate p<0.001 and p<0.01, respectively,against high fat diet; ( ) indicates the number of cases.

These results shows that the compounds according to the presentinvention are effective in controlling glycaemia in fasting conditions.This is an important aspect in the treatment of diabetes, whereinhepatic gluconeogenesis occurs during fasting periods (i.e. nocturnalrest).

The effect of CPT Inhibitors on Myocardial Ischemia

The compounds of the present invention are also effective in thetreatment of ischemia, in particular myocardial ischemia.

To this end, male Wistar rats, weighing 200-225 g, provided byCharles-River, were kept at constant temperature of 23°+/−1° C.,50+/−10% relative humidity, 12 hours light-dark cycle, fed with pellet4RF2 1 (Mucedola) tap water ad libitum.

The animals were anaesthetised with sodium Pentobarbital at the dose of70 mg/Kg i.p. Hearts were rapidly removed and put in a coldKrebs-Henseleit solution, before incannulation of aorta e subsequentperfusion according to Langendorff technique at 37° C. with a pressureof 100 cm water.

Perfusion medium (Krebs-Henseleit) at pH 7.4 consists in: 128 mM NaCl,4.7 mM KCl, 1 mM MgCl2, 0.4 mM Na2HPO4, 20.2 mM NaHCO3, 1.3 mM CaCl2, 5mM glucose. The medium was constantly oxygenated with carbogen (95% O2,5% CO2).

After a 10 min “conditioning” period, hearts were perfused in arecirculant apparatus for 20 min. with the same medium containing 0.6 mMpalmitate complexed with albumine (fraction V, fatty acid free), with orwithout the CPT inhibitor according to the present invention. By way ofexample ST 1364 was used at concentrations of 1 and 5 μM. After such aperiod ischemia was induced by reducing perfusion hydrostatic pressurefrom 100 cm to 20 cm for a period of 30 min. Reperfusion was startedre-establishing the starting pressure conditions (100 cm).Hearts werecontrolled for 20 min. the inhibitor is present also during reperfusionphase.

Lactate dehydrogenases (LDH) release was monitored in the effluent innormal oxygenation conditions, during ischemia, with a withdrawal ofmedium at 30′, and during reperfusion, with withdrawals at 1.5, 10, 15and 20 minutes.

LDH release in the effluent is remarkably reduced, during reperfusionresults significantly reduced in the presence of ST1364 at the dose of 5μM (FIG. 1). This result indicates a lower entity of cellular damagefrom reperfusion of the treated with respect to the controls.

Statistical analysis was carried out with Student's “t” test fornon-paired data.

The number of the cases for each group is six (n=6).

The following Table 5 reports the results.

TABLE 5 LDH release in perfusate (mU/ml/min) ST1364 ST1364 Control 1μM * 5 μM ** Basal 280 275 220 Ischemia 30′ 200 220 200 Reperfusion 1′640 480 410 Reperfusion 5′ 660 500 380 Reperfusion 10′ 670 495 380Reperfusion 15′ 700 510 320 Reperfusion 20′ 720 580 325

Statistical analysis was carried out with Student's “t” test fornon-paired data. * p<0.05 vs controls; **p<0.01 vs controls.

The number of the cases for each group is six (n=6).

LDH release in the effluent is remarkably reduced, during reperfusionresults significantly reduced in the presence of ST1364 at the dose of 5μM (FIG. 1). This result indicates a lower entity of cellular damagefrom reperfusion of the treated with respect to the controls.

In another aspect, the present invention provides a combination of atleast a compound of formula (I) with at least another active ingredientsuitable for the treatment of the disease of interest.

In the treatment or prevention of diabetes, the present inventionprovides a compound of formula (I), optionally in combination with asuitable well-known active ingredient, such as for example asulfonylurea, L-carnitine, fibrate and other agonists of peroxisomalproliferator activated receptor (PPAR-α), agonists of 9-cis retinoicacid activated receptor, such as RXR, in particular α-,β- andγ-isoforms, HMGCoA reductase inhibitor, β-sitosterol inhibitor,cholesterol acyltransferase inhibitor, biguanides, cholestyramine,angiotensin II antagonist, melinamide, nicotinic acid, fibrinogenreceptor antagonists, aspirin, α-glucosidase inhibitors, insulinsecretogogue, insulin and glucagon-like peptides (incretins) andagonists of PPAR-γ (such as thiazolidinediones or others).

In the treatment or prevention of obesity, the present inventionprovides a compound of formula (I), optionally in combination with ansuitable well-known active ingredient, such as for example fenfluramine,dexfenfluramine, phentiramine, a β-3-adrenergic receptor agonist.

In the treatment or prevention of high triglyceridhemia, the presentinvention provides a compound of formula (I), optionally in combinationwith an suitable well-known active ingredient.

The compounds according to the present invention are also useful in thetreatment or prevention of high cholesterol levels and in modulating HDLplasma levels, thus resulting beneficial in the treatment or preventionof the diseases related with these altered plasma levels. Examples ofrelated diseases are hypertension, obesity, atherosclerosis, diabetesand related conditions. The medicaments containing at least a compoundof the present invention may contain in combination at least anotheractive ingredient effective in the treatment or prevention of the abovementioned diseases. Examples of other active ingredient are fibrates,such as clofibrate, bezafibrate and gemfibrozil and other PPAR-αagonists; inhibitors of cholesterol biosynthesis, such as HMG-CoAreductase inhibitors, such as statins, namely lovastatin, simvastatinand pravastatin; inhibitors of cholesterol absorption for examplebeta-sitosterol and (acyl CoA:cholesterol acyltransferase) inhibitorsfor example melinamide; anion exchange resins for examplecholestyramine, colestipol or a dialkylaminoalkyl derivatives of across-linked dextran; nicotinyl alcohol, nicotinic acid or a saltthereof; vitamin E; thyromimetics and L-carnitine.

The compounds of the present invention may be orally administered in theform of a pharmaceutical composition, comprising a therapeuticallyeffective amount of at least a compound of formula (I) in admixture witha pharmaceutically acceptable vehicle and/or excipient. Examples of oralpharmaceutical compositions are hard or soft capsules, tablets,including sublingual administration, ampoules, sachets, elixirs,suspensions, syrups, and the like. Alternatively, the active ingredientsaccording to the present invention may be incorporated directly with thefood of the diet. The amount of active compound in such therapeuticallyuseful compositions is such that an effective dosage will be obtained.The active compounds can also be administered intranasally as, forexample, liquid drops or spray.

The tablets, pills, capsules, and the like may also contain a bindersuch as gum tragacanth, acacia, corn starch or gelatin; excipients suchas dicalcium phosphate; a disintegrating agent such as corn starch,potato starch, alginic acid; a lubricant such as magnesium stearate; anda sweetening agent such as sucrose, lactose or saccharin. When a dosageunit form is a capsule, it may contain, in addition to materials of theabove type, a liquid carrier such as a fatty oil.

Various other materials may be present as coatings or to modify thephysical form of the dosage unit. For instance, tablets may be coatedwith shellac, sugar or both. A syrup or elixir may contain, in additionto the active ingredient, sucrose as a sweetening agent, methyl andpropylparabens as preservatives, a dye and a flavoring such as cherry ororange flavor.

These active compounds may also be administered parenterally. Solutionsor suspensions of these active compounds can be prepared in pyrogen-freewater.

The pharmaceutical forms suitable for injectable use include sterileaqueous solutions or dispersions and sterile powders for theextemporaneous preparation of sterile injectable solutions ordispersions.

If desired, or deemed necessary, the pharmaceutical compositions may bein the controlled- release form. Various techniques for preparing theseforms are known.

General reference for pharmaceutical compositions can be made to“Remington's Pharmaceutical Sciences Handbook”, Mack Pub. N.Y. USA.

The effective dosage of active ingredient employed may vary depending onthe particular compound employed, the mode of administration, thecondition being treated and the severity of the condition being treated.

The compositions are formulated and administered in the same generalmanner as detailed below. The compounds of the present invention can beused effectively alone or in combination with one or more additionalactive agents depending on the desired target therapy. Combinationtherapy includes administration of a single pharmaceutical dosageformulation which contains a compound of formula I and one or moreadditional active agents, as well as administration of a compound offormula I and each active agent in its own separate pharmaceuticaldosage formulation. For example, a compound of formula I and an HMG-CoAreductase inhibitor can be administered to the patient together in asingle oral dosage composition such as a tablet or capsule, or eachagent administered in separate oral dosage formulations. Where separatedosage formulations are used, a compound of formula I and one or moreadditional active agents can be administered at essentially the sametime, i.e., concurrently, or sequentially; combination therapy isunderstood to include all these regimens.

An example of combination treatment or prevention of atherosclerosis iswherein a compound of formula I is administered in combination with oneor more of the following active agents: an antihyperlipidemic agent; aplasma HDL-raising agent; an antihypercholesterolemic agent such as acholesterol biosynthesis inhibitor, for example an HMG-CoA reductaseinhibitor, an HMG-CoA synthase inhibitor, a squalene epoxidaseinhibitor, or a squalene synthetase inhibitor (also known as squalenesynthase inhibitor); an acyl-coenzyme A: cholesterol acyltransferase(ACAT) inhibitor such as melinamide; probucol; nicotinic acid and thesalts thereof and niacinamide; a cholesterol absorption inhibitor suchas beta-sitosterol; a bile acid sequestrant anion exchange resin such ascholestyramine, colestipol or dialkylaminoalkyl derivatives of across-linked dextran; an LDL (low density lipoprotein) receptor inducer;fibrates such as clofibrate, bezafibrate, fenofibrate, and gemfibroziland other PPAR-α agonists, L-carnitine; vitamin B₆ and thepharmaceutically acceptable salts thereof; vitamin B₁₂; anti-oxidantvitamins such as vitamin C and E and beta carotene; a beta-blocker; anangiotensin II antagonist; an angiotensin converting enzyme inhibitor;and a platelet aggregation inhibitor such as fibrinogen receptorantagonists (i.e., glycoprotein IIb/IIIa fibrinogen receptorantagonists) and aspirin. The compounds of formula I can be administeredin combination with more than one additional active agent.

Another example of combination therapy can be seen in treating obesityor obesity-related disorders, wherein-the compounds of formula I may beeffectively used in combination with for example, fenfluramine,dexfenfluramine, phentiramine and β-3 adrenergic receptor agonist agentsand L-carnitine.

Another example of combination therapy can be seen in treating diabetesand related disorders wherein the compounds of formula I can beeffectively used in combination with for example sulfonylureas,biguanides, α-glucosidase inhibitors, other insulin secretogogues,insulin and glucagon-like peptides (incretins) and agonists of PPAR-γ(such as thiazolidinediones or others) as well as the active agentsdiscussed above for treating atherosclerosis.

What is claimed is:
 1. A compound of formula (I)

wherein: X⁺ is N⁺(R₁,R₂,R₃), wherein R₁,R₂,R₃, being the same ordifferent, are selected in the group consisting of hydrogen, a C₁-C₉straight or branched alkyl group, —CH═NH(NH₂), —NH₂, and —OH; or one ormore R₁, R₂ and R₃, together with the nitrogen atom which they arelinked to, form a saturated or unsaturated, monocyclic or bicyclicheterocyclic system; with the proviso that at least one of the R₁, R₂and R₃ is different from hydrogen; Z is selected from —OR₄, —OCOOR₄,—OCONHR₄, —OCSNHR₄, —OCSOR₄, —NHR₄, —NHCOR₄, —NHCSR₄, —NHCOOR₄,—NHCSOR₄, —NHCONHR₄, —NHCSNHR₄, —NHSOR₄, —NHSONHR₄, —NHSO₂R₄,—NHSO₂NHR₄, and —SR₄, wherein —R₄ is a C₁-C₂₀ saturated or unsaturated,straight or branched alkyl group, optionally substituted with an A₁group, wherein A₁ is selected from the group consisting of a halogenatom, or an aryl, heteroaryl, aryloxy or heteroaryloxy group, said aryl,heteroaryl, aryloxy or heteroaryloxy groups being optionally substitutedwith one or more C₁-C₂₀ saturated or unsaturated, straight or branchedalkyl or alkoxy group and/or halogen atom; Y⁻ is selected from the groupconsisting of —COO⁻, PO₃H⁻, —OPO₃H⁻, tetrazolate-5-yl; with the provisothat when Z is —NHCOR₄, Y is —COO⁻, then R₄ is C₂₀ alkyl; with theproviso that when Z is —NHSO₂R₄, Y⁻ is —COO⁻, then R₄ is not tolyl; withthe proviso that when Z is —NHR₄, X⁺ is trimethylammonium and Y⁻ is—COO⁻, then R₄ is not C₁-C₆ alkyl, their (R,S) racemic mixtures, theirsingle R or S enantiomers, or their pharmaceutically acceptable salts.2. A compounds according to claim 1, wherein R₁, R₂ and R₃ are methyl.3. A compounds according to claim 1, wherein the heterocyclic systemformed by R₁, R₂ and R₃ together with nitrogen is selected from thegroup consisting of morpholinium, quinuclidinium, pyridinium,quinolinium and pyrrolidinium.
 4. A compound according to claim 1,wherein R₁ and R₂ are H, R₃ is selected from the group consisting of—CH═NH(NH₂), —NH₂ and —OH.
 5. A compound according to claim 1, wherein Zis selected from the group consisting of ureido (—NHCONHR₄) or carbamate(—OCONHR₄), and R₄ is a C₇-C₂₀ saturated or unsaturated, straight orbranched alkyl group.
 6. A compound according to claim 5, wherein R₄ isa C₉-C₁₈ saturated or unsaturated, straight or branched alkyl group. 7.A compound selected from the group consisting ofR,S-4-trimethylammonium-3-(nonylcarbamoyl)-aminobutyrate;R,S-4-quinuclidinium-3-(tetradecyloxycarbonyl)-oxybutyrate;R,S-4-trimethylammonium-3-(nonylcarbamoyl)-oxybutyrate;R,S-4-trimethylammonium-3-(nonyloxycarbonyl)-oxybutyric acid chloride;R,S-4-trimethylphosphonium-3-(nonylcarbamoyl)-oxybutyrate;R,S-4-trimethylammonium-3-(octyloxycarbonyl)-aminobutyrate;R,S-4-trimethylammonium-3-(nonyloxycarbonyl)-aminobutyrate;R,S-4-trimethylammonium-3-octyloxybutyrate;R,S-4-trimethylammonium-3-tetradecyloxybutyrate;R,S-1-guanidinium-2-tetradecyloxy-3-(tetrazolate-5-yl)-propane;R,S-4-trimethylammonium-2-tetradecyloxy-3-(tetrazolate-5-yl)-propane;R,S-3-quinuclidium-2-(tetradecyloxycarbonyl)-oxy-1-propanephosphonatemonobasic;R,S-3-trimethylammonium-2-(nonylaminocarbonyl)-oxy-1-propanephosphonatemonobasic;R,S-3-pyridinium-2-(nonylaminocarbonyl)-oxy-1-propanephosphonic acidchloride; R-4-trimethylammonium-3-(tetradecylcarbamoyl)-aminobutyrate;R-4-trimethylammonium-3-(undecylcarbamoyl)-aminobutyrate;R-4-trimethylammonium-3-(heptylcarbamoyl)-aminobutyrate;R,S-4-trimethylammonium-3-(nonylthiocarbamoyl)-aminobutyrate;R-4-trimethylammonium-3-(nonylcarbamoyl)-aminobutyrate;S-4-trimethylammonium-3-(nonylcarbamoyl)-aminobutyrate;S-4-trimethylammonium-3-(tetradecylcarbamoyl)-aminobutyrate;R,S-4-trimethylammonium-3-tetradecylaminobutyrate;R,S-4-trimethylammonium-3-octylaminobutyrate;R,S-4-trimethylammonium-3-(decansulfonyl)aminobutyrate;R,S-4-trimethylammonium-3-(nonylsulfamoyl)aminobutyrate;S-4-trimethylammonium-3-(dodecansulfonyl)aminobutyrate;R-4-trimethylammonium-3-(dodecansulfonyl)aminobutyrate;S-4-trimethylammonium-3-(undecylsulfamoyl)aminobutyrate;R-4-trimethylammonium-3-(undecylsulfamoyl)aminobutyrate;R-4-trimethylammonium-3-(dodecylcarbamoyl)aminobutyrate;R-4-trimethylammonium-3-(10-phenoxydecylcarbamoyl)aminobutyrate; andR-4-trimethylammonium-3-(trans-β-styrenesulfonyl)aminobutyrate.
 8. Aprocess for the preparation of a compound of claim 1, wherein Z iscarbonate (—OCOOR₄), carbamate (—NHCOOR₄), thiocarbamate (—OCSNHR₄) orthiocarbonate (—OCSOR₄), said process comprising reactingX+—CH₂—CH(OH)—CH₂—Y—, of the desired structure, optionally protected onthe acid Y— group, respectively with an alkyl chloroformate, alkylisocyanate, alkyl isothiocyanate or alkyl thiochloroformate, wherein thealkyl moiety is the desired R₄ alkyl group, to produce the desiredcompound.
 9. A process for a preparation of a compound of claim 1,wherein Z is amide (—NHCOR₄), thioamide (—NHCSR₄), carbamate (—NHCOOR₄),thiocarbamate (—NHCSOR₄), ureido (—NHCONHR₄), thioureido (—NHCSNHR₄),sulfinamide (—NHSOR₄), sulfonamide (—NHSO₂R₄), sulfinamoylamino(—NHSONHR₄), and sulfamide (—NHSO₂NHR₄), said process comprisingreacting X⁺—CH₂—CH(OH)—CH₂—Y⁻, of the desired structure, optionallyprotected on the acid Y⁻ group, respectively with an acyl chloride,thioacyl chloride, alkyl chloroformate, alkyl thiochloroformate, alkylisocyanate, alkyl thioisocyanate, alkyl sulfinyl chlorides, alkylsulfonyl chlorides, SOCl₂ and alkyl amines, alkyl sulfamoyl chloride orSOCl₂ and alkyl amine, wherein the alkyl moiety is the desired R₄ alkylgroup, to produce the desired compound.
 10. A process for thepreparation of a compound of claim 1, wherein Z is —OR₄ or —SR₄, saidprocess comprising the steps of: (a) reacting a carbonyl compound offormula Hal-CH₂—CO—CH₂—COOR′, wherein Hal is a halogen atom and R′ isthe residue of a suitable ester, with respectively alcohols and thiolsR₄OH or R₄SH, to give the respective ketal or thioketal; (b)transforming the respective ketal or thioketal into the respective etheror thioether; (c) substituting the Hal atom with an azido group, and (d)transforming the azido group into the X+ group to produce the desiredcompound.
 11. A process for the preparation of a compound of claim 1,wherein Z is —NHR₄, said process comprising reacting ofX⁺—CH₂—CH(NH₂)—CH₂—Y⁻ of the desired structure, optionally protected onthe acid Y⁻ group, with alkane carbaldheydes, wherein the alkyl moietyis a one-term lower homologue of the desired R₄, and subsequentreduction, to produce the desired compound.
 12. A pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundof claim 1, in admixture with a pharmaceutically acceptable vehicle orand excipient.
 13. The pharmaceutical composition according to claim 12,wherein an active ingredient suitable for the treatment of diabetes isalso present and is selected from the group consisting of sulfonylurea,L-carnitine, fibrate and other agonists of peroxisomal proliferatoractivated receptor (PPAR-α), HMG-CoA reductase inhibitor, β-sitosterolinhibitor, cholesterol acyltransferase inhibitor, biguanides,cholestyramine, angiotensin II antagonist, melinamide, nicotinic acid,fibrinogen receptor antagonists, aspirin, α-glucosidase inhibitors,insulin secretogogue, insulin and glucagon-like peptides and agonists ofPPAR-γ.
 14. A pharmaceutical composition according to claim 12, alsoincluding an active ingredient suitable for the treatment of obesityselected from the group consisting of fenfluramine, dexfenfluramine,phentiramine, and a β-3-adrenergic receptor agonist.
 15. Apharmaceutical composition according to claim 12, also including anactive ingredient suitable for the treatment of high cholesterol levelsand in modulating HDL plasma levels, which is selected from the groupconsisting of fibrates, and other PPAR-α agonists; inhibitors ofcholesterol biosynthesis, HMG-CoA reductase inhibitors, statins,inhibitors of cholesterol absorption, acyl CoA:cholesterolacyltransferase inhibitors, anion exchange resins, nicotinyl alcohol,nicotinic acid or a salt thereof, vitamin E, thyromimetics andL-carnitine.
 16. A method for treating a subject having hyperactivecarnitine palmitoyl-transferase comprising administering to said subjectan effective amount of a compound of claim
 1. 17. A method for treatinga subject having hyperglycaemia, diabetes, heart failure or ischemiacomprising administering to said subject an effective amount of acompound of claim
 1. 18. A method for treating a subject having obesitycomprising administering to said subject an effective amount of acompound of claim
 1. 19. A method for treating a subject having hightriglyceridemia comprising administering to said subject an effectiveamount of a compound of claim
 1. 20. A method for treating a subjecthaving hypertension comprising administering to said subject aneffective amount of a compound of claim
 1. 21. A method of modulatinghigh cholesterol levels or MDL plasma levels in a subject in need ofsame, said method comprising administering to said subject an effectiveamount of a compound of claim 1.